Crystal structure of the disulfide-stabilized Fv fragment of anticancer antibody B1: Conformational influence of an engineered...
TY - JOUR. T1 - Crystal structure of the disulfide-stabilized Fv fragment of anticancer antibody B1. T2 - Conformational influence of an engineered disulfide bond. AU - Almog, Orna. AU - Benhar, Itai. AU - Vasmatzis, George. AU - Tordova, Maria. AU - Lee, Byungkook. AU - Pastan, Ira. AU - Gilliland, Gary L.. PY - 1998/5/1. Y1 - 1998/5/1. N2 - A recombinant Fv construct of the B1 monoclonal antibody that recognizes the Lewis(Y)-related carbohydrate epitope on human carcinoma cells has been prepared. The Fv is composed of the polypeptide chains of the V(H) and V(L) domains expressed independently and isolated as inclusion bodies. The Fv is prepared by combining and refolding equimolar amounts of guanidine chloride solubilized inclusion bodies. The Fv is stabilized by an engineered interchain disulfide bridge between residues V(L)100 and V(H)44. This construct has a similar binding affinity as that of the single-chain construct (Benhar and Pastan, Clin. Cancer Res. 1:1023-1029, 1995). The B1 ...
Escherichia coli Skp chaperone coexpression improves solubility and phage display of single-chain antibody fragments<...
TY - JOUR. T1 - Escherichia coli Skp chaperone coexpression improves solubility and phage display of single-chain antibody fragments. AU - Hayhurst, Andrew. AU - Harris, William J.. PY - 1999/4. Y1 - 1999/4. N2 - Expression of single-chain antibody fragments (scAb) in the periplasm of Escherichia coli often results in low soluble product yield and cell lysis. We have increased scab solubility and prevented cell culture lysis by coexpressing the E. coli Skp chaperone gene. A mutant Skp cistron was linked to a bacteriophage T7 gene 10 translational initiation region and placed either downstream of a scab gene within an isopropyl β-D-thiogalactopyranoside-inducible expression cassette or on a separate colE1-compatible arabinose-inducible vector. Increases in scab solubility reflected the amount of coexpressed Skp. A bacteriophage display vector that was also engineered to coexpress Skp permitted display of a virtually undisplayable scab and should prove useful in expanding library sizes.. AB - ...
Production and characterisation of novel recombinant antibody fragments against sialic acid - DORAS - DCU
Sialic acids are a family of acidic monosaccharides that typically reside as terminal moieties on N- and O-linked glycans. These sugars are actively involved in a plethora of biological phenomena, ranging from cell-cell adhesion and recognition, intracellular signalling events, pathogen attack, viral infection and inflammatory disease. In addition, many cancer-related antigens contain terminal sialic acids or altered sialylation patterns. The identification of sialic acid-specific antibodies is important in the fields of basic research and diagnostics. Therefore the primary objective of this thesis was the generation and characterisation of recombinant antisialic acid antibodies.A single-chain antibody fragment (scFv) library was constructed from a chicken that was immunised with a novel synthetic sialic acid protein-conjugate (Neu5Gchumanserum albumin). The scFv library was biopanned using a second novel sialoneoglycoconjugate(Neu5Gc-bovine serum albumin). Anti-sialic scFvs were isolated ...
Recombinant Anti-Human CD4 Antibody scFv Fragment - Creative Biolabs
Creative Biolabs can provide Recombinant Anti-Human CD4 Antibody scFv Fragment of scFv Fragment from Humanized (from mouse) IgG1 - kappa
Recombinant Anti-Human IL6 Antibody scFv Fragment - Creative Biolabs
Creative Biolabs can provide Recombinant Anti-Human IL6 Antibody scFv Fragment of scFv Fragment from Humanized (from rat) IgG4 - kappa
British Library EThOS: Constructions and expressions of anti-ABA scFv genes in bacteria, yeast and plants
Three anti-ABA single-chain Fv (scFv) antibody genes, namely ABA26, MAC61, MAC252 scFvs, had been constructed from mouse and rat hybridomas and expressed in bacteria, yeast and plants. All of these scFv genes could be expressed in E. coli using the T7 promoter, either targeted to the E. coli periplasm or cytoplasm, albeit at comparatively low levels. The cytoplasmically located scFv proteins were in the form of insoluble fraction and did not therefore exhibit any binding activities to the ABA. The majority of the periplasmically located scFv proteins were retained in the bacterial cytoplasm as insoluble bodies with the ompA signal peptide attached to them. Nevertheless, the positive signal in ELISA test indicated that a small portion of scFv proteins were in the form of soluble functional scFv proteins. All the three periplasmically expressed scFv proteins had specific ABA binding activities. However, the affinity constants of the scFv proteins were found to be 5 to 10 fold lower than those of ...
Modulation of the CD40-CD40 ligand interaction using human anti-CD40 single-chain antibody fragments obtained from the n-CoDeR...
article{c58957be-5df1-4efc-b433-f02e50dc2d3f, abstract = {CD40 plays a central regulatory role in the immune system and antibodies able to modulate CD40 signalling may consequently have a potential in immunotherapy, in particular for treatment of lymphomas and autoimmune disease like multiple sclerosis. As a first step to achieve this goal, we describe the selection and characterization of a novel set of fully human anti-CD40 antibody fragments (scFv) from a phage display library (n-CoDeR). In order to determine their biological potential, these antibody fragments have been analysed for their ability to promote B-cell activation, rescue from apoptosis and to block the CD40-CD40 ligand (CD40L) interaction. The selected cohort of human scFv could be subcategorized, each expressing a distinct functional signature. Thus scFv were generated that induced B-cell proliferation, rescued B cells from apoptosis and blocked the CD40-CD40L interaction to different extents. In particular, one of the scFv ...
Antibody fragments: Prolonging circulation half-life special issue-antibody research
Antibodies are currently the fastest growing class of
therapeutic proteins. When antibody fragments are included, there are over
thirty-five antibody-based medicines approved for human therapy. Many more
antibody and antibody-like fragments are being evaluated clinically.
Production of antibody fragments can be efficient and their compact size can
allows for better tissue extravasation into solid tumors than full antibodies.
Unfortunately, a key limitation of antibody fragments for systemic use is
their short half-life in circulation. Prolonging their circulation half-life
can be accomplished clinically by the covalent conjugation of the antibody
fragment to the water-soluble polymer, poly(ethylene glycol) (PEG). Many polymers
and strategies are also being pursued to increase antibody fragment half-life.
Sequence Similarity - 1F3R: COMPLEX BETWEEN FV ANTIBODY FRAGMENT AND AN ANALOGUE OF THE MAIN IMMUNOGENIC REGION OF...
1F3R: The third-dimensional structure of the complex between an Fv antibody fragment and an analogue of the main immunogenic region of the acetylcholine receptor: a combined two-dimensional NMR, homology, and molecular modeling approach.
RCSB PDB - 1F3R: COMPLEX BETWEEN FV ANTIBODY FRAGMENT AND AN ANALOGUE OF THE MAIN IMMUNOGENIC REGION OF THE...
1F3R: The third-dimensional structure of the complex between an Fv antibody fragment and an analogue of the main immunogenic region of the acetylcholine receptor: a combined two-dimensional NMR, homology, and molecular modeling approach.
Murugesan Subbarayan | Molecular Imaging Program at Stanford | Stanford Medicine
Kenanova V, Olafsen T, Crow DM, Sundaresan G, Subbarayan M, Carter NH, Ikle DN, Yazaki PJ, Chatziioannou AF, Gambhir SS, Williams LE, Shively JE, Colcher D, Raubitschek AA, Wu AM. Tailoring the pharmacokinetics and positron emission tomography imaging properties of anti-carcinoembryonic antigen single-chain Fv-Fc antibody fragments. Cancer Res. 2005;65(2):622-31 ...
e-space
BACKGROUND: Monoclonal antibodies and antibody fragments are powerful biotherapeutics for various debilitating diseases. However, high production costs, functional limitations such as inadequate pharmacokinetics and tissue accessibility are the current principal disadvantages for broadening their use in clinic. METHODOLOGY AND PRINCIPAL FINDINGS: We report a novel method for the long-term delivery of antibody fragments. We designed an allogenous immunoisolated implant consisting of polymer encapsulated myoblasts engineered to chronically release scFv antibodies targeted against the N-terminus of the Aβ peptide. Following a 6-month intracerebral therapy we observed a significant reduction of the production and aggregation of the Aβ peptide in the APP23 transgenic mouse model of Alzheimers disease. In addition, functional assessment showed prevention of behavioral deficits related to anxiety and memory traits. CONCLUSIONS AND SIGNIFICANCE: The chronic local release of antibodies using ...
Biomimic Discovery
The biomimc development team is using both phage-display and SELEX techniques to develop (i) glycan specific antibody molecules and (ii) synthetic mimics of natural lectins to improve specificity to individual glycan epitopes. These mimics are in the form of either recombinant antibody fragments or as DNA and RNA aptamers. Increased specificity to glycan targets is essential in developing new diagnostic reagents and also in the analysis of targeted glycans present on recombinant biopharmaceuticals for bioprocessing QA and regulatory control. Once new biomimics are identified they are utilised and developed as both stand along bioassays and also incorporated into our custom array platforms, thereby increasing array reagent repertoire, broadening specificity, increasing signature information and providing additional HTP analysis markers.. Utilising our array and biomimic technologies we continue to advance the glyco-profiling cell surface to identify key changes that occur during disease ...
CD45RA Antibody, anti-human, REAlease® - Releasable antibodies - MACS Antibodies - Products - Miltenyi Biotec - Belgique
Clone REAL164 is an antibody fragment derived from the full CD45RA antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity. Clone REAL164 recognizes the human CD45RA antigen which is expressed on naive CD4+ and CD8+ T cells as well as on CD8+ effector T cells. CD45RA is also present on subsets of B and NK cells and on plasmacytoid dendritic cells. The CD45RA antibody recognizes the 220 kDa isoform of the leukocyte common antigen (LCA), a transmembrane tyrosine phosphatase. The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes. - Belgique
CD54 (ICAM-1) Antibody, anti-human, REAlease® - Releasable antibodies - MACS Antibodies - Products - Miltenyi Biotec -...
Clone REAL146 is an antibody fragment derived from the full CD54 (ICAM-1) antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.Clone REAL146 recognizes human CD54, a 85-110 kDa type I transmembrane glycoprotein, which also known as intercellular adhesion molecule 1 (ICAM-1). CD54 is continuously present in low concentrations in the membranes of monocytes/macrophages, lymphocytes, activated endothelial cells, granulocytes, and dendritic cells. Its expression is regulated by inflammatory cytokines and can be induced by interleukin-1 and tumor necrosis factor. CD54 is a ligand for LFA-1, a receptor found on leukocytes. When activated, leukocytes bind to endothelial cells via CD54/LFA-1 and then transmigrate into tissues. CD54 has been shown to interact with CD11a, EZR, and CD18. More recently, CD54 has been characterized as a site for the cellular entry of human rhinovirus
US Patent # 9,365,636. Human cytomegalovirus neutralizing antibodies and use thereof - Patents.com
The invention relates to neutralizing antibodies, and antibody fragments thereof, having high potency in neutralizing hCMV, wherein said antibodies and antibody fragments are specific for one, or a c
Novel Small Antibody Fragment: A Valuable Tool in Crystallography - ResOU
Achieving quality diffracting crystals remains the major bottleneck in macromolecular x-ray crystallography. Antibody fragments have been used to aid structural analysis of complex macromolecules that are otherwise crystallization-resistant. However, there are issues such as stability, compatibility, and cost. Osaka University researchers succeeded in creating a novel antibody fragment termed
UVA Licensing & Ventures Group
locations, including much of West Africa. The World Health Organization has called for new detection kits that are cheaper, provide readout in less than 30 minutes and require no more than two simple steps.. The science to support such a kit is complex, and thats what U.Va. researchers are trying to tease out.. Such kits may detect virus antigens in blood samples, Derewenda said. However, the monoclonal antibodies needed for this method are relatively expensive and not easy to obtain using traditional technology. We are pursuing an alternative strategy, in which synthetic antibody fragments are engineered to recognize a different target in the nucleoprotein.. Derewenda and his team will be able to move forward more quickly thanks to $80,000 in funding from the Ivy Foundation Biomedical Innovation Grants ...
Intrabodies based on intracellular capture frameworks that bind the RAS protein with high affinity and impair oncogenic...
The most important requirements for intracellular antibodies as therapeutic or bioscience research tools is that these antibodies (or antibody fragments) exhibit good expression levels and are functional within any compartment of mammalian cells. Ideally, these also work as functional reagents for in vitro use. There are severe limitations, and few scFv fragments derived from hybridomas are stable under a reducing environment without modification, even if they have good affinity in vitro. The IAC technology described here, and previously (Tse et al., 2002b; Visintin et al., 2002), overcomes these difficulties as it is based on an in vivo genetic screen for the direct isolation of functional scFvs.. The IAC approach has several advantages compared with other screening methods. It is based on the yeast two‐hybrid in vivo assay (Fields and Song, 1989), which works as direct cytoplasmic selection of scFvs (Visintin et al., 1999). In addition, it theoretically allows the selection of antibody ...
Involving immunoglobulin or antibody fragment (e.g., f(ab`)2, fab`, fv, fc, heavy chain, light chain, etc.) > Patents - Class:...
Name: Drug, bio-affecting and body treating compositions > Involving immunoglobulin or antibody fragment (e.g., f(ab`)2, fab`, fv, fc, heavy chain, light chain, etc ...
Technology Overview - affinity BIO
Affinity BIO has successfully employed its ReD3 screening system against a range of oncology and viral pMHC targets, generating single-chain antibodies (scFv) that have both high-affinity AND high-specificity to a single pMHC. The ability of the ReD3 system to do this can be explained by a combination of attributes that are significantly differentiated from standard antibody discovery technologies such as phage display.. The ability to target intra-cellular proteins creates a host of new therapeutic options, potentially enabling new treatments of various forms of cancer, viral infections and autoimmune diseases.. ...
An Industrial Platform Solution for Antibody Fragment PurificationBioProcess International
A recombinant technology based on protein L, that enables a cost-effective affinity chromatography platform approach for Fc-lacking antibody fragments such as Fabs and dAbs.
CMU Intellectual Repository: Production of single-chain fragment variable (scFv) antibodies specific to plasma membrane...
2020 Taylor & Francis Group, LLC. Distinguishing between bull Y- and X-bearing sperm populations is advantageous for techniques using sexed bull semen. The aim of this study was to produce a single-chain fragment variable (scFv) antibody against plasma membrane epitopes on bull Y-bearing sperm. Variable heavy (VH)- and variable light (VL)-region genes generated from a hybridoma cell secreting a specific Y-bearing sperm monoclonal antibody (mAb-1F9) were cloned and expressed. The expected sizes of the DNA bands were ∼350 bp for the VH gene and ∼318 bp for the VL gene. The VH and VL genes were generated and used to construct an scFv gene (∼650 bp), which was expressed in E.coli TG1 cells and produced the corresponding soluble scFv antibody. Compared with the parent mAb-1F9, the scFv antibodies presented a high affinity for Y-bearing sperm and low cross-reactivity with X-bearing sperm. An immunofluorescence analysis confirmed that the scFv antibodies and mAb-1F9 recognize epitopes on the ...
Development and Characterization of Three Recombinant Single Chain Antibody Fragments (scFvs) Directed against the CD19 Antigen...
Antibodies that recognize antigens restricted to leukemia, lymphoma, and normal hematopoietic cells represent a unique opportunity to develop therapeutics, because they have the potential for relatively selective treatment of these diseases. Antibodies that recognize the CD19 antigen found on normal and malignant B cells, but not on stem cells, have been used to develop immunoconjugates. However, these conjugates are large and might be suboptimal in tumor penetration when compared to molecules using smaller single chain Fv (scFv) antibody fragments. scFv has the advantage of being a molecularly engineered homogeneous molecule. In this report, we demonstrate the cloning, expression, and binding of three anti-CD19 antibodies as scFvs. All three scFvs were successfully cloned and expressed. FVS191, derived from cell line B43, and FVS192, derived from SJ25C1, were properly refolded and bound CD19 antigen in FACS competition assays. These anti-CD19 scFv should be useful in the further development of ...
A large semi-synthetic single-chain Fv phage display library based on chicken immunoglobulin genes | BMC Biotechnology | Full...
Antibody fragments selected from large combinatorial libraries have numerous applications in diagnosis and therapy. Most existing antibody repertoires are derived from human immunoglobulin genes. Genes from other species can, however, also be used. Because of the way in which gene conversion introduces diversity, the naïve antibody repertoire of the chicken can easily be accessed using only two sets of primers. With in vitro diagnostic applications in mind, we have constructed a large library of recombinant filamentous bacteriophages displaying single chain antibody fragments derived from combinatorial pairings of chicken variable heavy and light chains. Synthetically randomised complementarity determining regions are included in some of the heavy chains. Single chain antibody fragments that recognise haptens, proteins and virus particles were selected from this repertoire. Affinities of three different antibody fragments were determined using surface plasmon resonance. Two were in the low nanomolar
In Vivo Detection of Amyloid-\(\beta\) Deposits Using Heavy Chain Antibody Fragments in a Transgenic Mouse Model for Alzheimer...
This study investigated the in vivo properties of two heavy chain antibody fragments (V\(_H\)H), ni3A and pa2H, to differentially detect vascular or parenchymal amyloid-\(\beta\) deposits characteristic for Alzheimers disease and cerebral amyloid angiopathy. Blood clearance and biodistribution including brain uptake were assessed by bolus injection of radiolabeled (V\(_H\)H) in APP/PS1 mice or wildtype littermates. In addition, in vivo specificity for A\(\beta\) was examined in more detail with fluorescently labeled (V\(_H\)H) by circumventing the blood-brain barrier via direct application or intracarotid co-injection with mannitol. All (V\(_H\)H) showed rapid renal clearance (10-20 min). Twenty-four hours post-injection \(^{99m}\)Tc-pa2H resulted in a small yet significant higher cerebral uptake in the APP/PS1 animals. No difference in brain uptake were observed for \(^{99m}\)Tc-ni3A or DTPA(\(^{111}\)In)-pa2H, which lacked additional peptide tags to investigate further clinical applicability. ...
Molecules | Free Full-Text | Production of Single-Chain Fv Antibodies Specific for GA-Pyridine, an Advanced Glycation End...
Due to their lower production cost compared with monoclonal antibodies, single-chain variable fragments (scFvs) have potential for use in several applications, such as for diagnosis and treatment of a range of diseases, and as sensor elements. However, the usefulness of scFvs is limited by inhomogeneity through the formation of dimers, trimers, and larger oligomers. The scFv protein is assumed to be in equilibrium between the closed and open states formed by assembly or disassembly of VH and VL domains. Therefore, the production of an scFv with equilibrium biased to the closed state would be critical to overcome the problem in inhomogeneity of scFv for industrial or therapeutic applications. In this study, we obtained scFv clones stable against GA-pyridine, an advanced glycation end-product (AGE), by using a combination of a phage display system and random mutagenesis. Executing the bio-panning at 37 °C markedly improved the stability of scFvs. We further evaluated the radius of gyration by small-angle
Expression of single-chain variable fragments fused with the Fc-region of rabbit IgG in Leishmania tarentolae | Microbial Cell...
Antibodies are applied in both basic research and diagnostics, and represent an increasingly important class of therapeutics. Monoclonal antibodies is the largest and fastest growing class of protein pharmaceuticals [1]. In the discovery and development of these antibodies, antibody fragments such as the antigen binding fragment (Fab), the single-chain variable fragment (scFv), and the single variable domains (VH and VL, collectively sdAb) are often employed [2]. The present recombinant antibody discovery platforms, such as ribosome and phage display [3], enable easy screening and selection of antibody fragments against virtually any antigen [4]. Based on the initial screening or selection, a number of candidate antibodies are obtained [3]. Often, these recombinant antibody candidates can be expressed in E. coli. However, although the field of recombinant protein expression in E. coli is developed and expanded [5, 6], the codon usage and folding dynamics of some recombinant antibody clones are ...
Go to Intracellular single-chain variable fragments directed to the Src homology 2 domains of Syk partially inhibit Fc epsilon...
Intracellular expression of Ab fragments has been efficiently used to inactivate therapeutic targets, oncogene products, and to induce viral resistance in plants. Ab fragments expressed in the appropriate cell compartment may also help to elucidate the functions of a protein of interest. We report in this study the successful targeting of the protein tyrosine kinase Syk in the RBL-2H3 rat basophilic leukemia cell line. We isolated from a phage display library human single-chain variable fragments (scFv) directed against the portion of Syk containing the Src homology 2 domains and the linker region that separates them. Among them, two scFv named G4G11 and G4E4 exhibited the best binding to Syk in vivo in a yeast two-hybrid selection system. Stable transfectants of RBL-2H3 cells expressing cytosolic G4G11 and G4E4 were established. Immunoprecipitation experiments showed that intracellular G4G11 and G4E4 bind to Syk, but do not inhibit the activation of Syk following FcepsilonRI aggregation, ...
Engineering AAV-2 Targeting Vectors: A New Insertion Site and scFv Driven Vectors - Kölner UniversitätsPublikationsServer
Gene therapy can be defined as the introduction of nucleic acids into cells with the purpose of altering the course of a medical condition or disease. In many clinical settings, efficient and successful gene therapy relies on the delivery of genes to specific cells in the human body. Such specific delivery can only be achieved through the design of vehicles/vectors that are able to recognize the target cell � targeting. Adeno-associated virus type 2, a non-pathogenic human virus, has received an increased amount of attention as a vector for gene therapy since it was first cloned into a bacterial plasmid in 1982. Although the first attempt to construct an AAV-2 targeting vector made use of a single chain antibody fragment, it was the insertion of small peptide ligands into the AAV-2 capsid that marked the beginning of AAV-2 vector targeting. Sequence alignment of AAV-2 with CPV led to the discovery of amino acid position 587. Several reports have shown that small peptide ligands, once inserted ...
Blood-Based Oligomeric and Other Protein Variant Biomarkers to Facilitate Pre-Symptomatic Diagnosis and Staging of Alzheimers...
TY - JOUR. T1 - Blood-Based Oligomeric and Other Protein Variant Biomarkers to Facilitate Pre-Symptomatic Diagnosis and Staging of Alzheimers Disease. AU - Williams, Stephanie M.. AU - Schulz, Philip. AU - Rosenberry, Terrone L.. AU - Caselli, Richard J.. AU - Sierks, Michael R.. PY - 2017. Y1 - 2017. N2 - Oligomeric forms of amyloid-β (Aβ), tau, and TDP-43 play important roles in Alzheimers disease (AD), and therefore are promising biomarkers. We previously generated single chain antibody fragments (scFvs) that selectively bind disease-related variants of these proteins including A4, C6T, and E1, which bind different oligomeric Aβ variants; D11C, which binds oligomeric tau; and AD-TDP1 and AD-TDP2, which bind disease related TDP-43 variants. To determine the utility of these disease-related variants as early biomarkers, we first analyzed 11 human sera samples obtained ∼2 years prior to an initial mild cognitive impairment (MCI) diagnosis. While the subsequent diagnoses for the cases ...
Molecular cloning of a monoclonal anti-tumor antibody specific for the Tn antigen and expression of an active single-chain Fv...
Link to Pubmed [PMID] - 9309422. Hybridoma 1997 Aug;16(4):317-24. We report here the first amino acid sequence of an anti-Tn monoclonal antibody raised against human breast cancer cells and show that a single chain Fv fragment of this IgM retains the Tn-binding specificity as defined by functional assays with asialo-OSM and membrane extracts from MCF-7 cells. Sequence comparisons and molecular modeling of 83D4 indicate that the antibody combining site displays a cavity-like feature primarily defined by the CDR H1 and H2 loops. This pocket could accommodate a single Tn molecule, thus, suggesting a structural explanation for the predominant expression of a particular VH gene segment in a group of antibodies that recognize tumor-associated antigens arising from an aberrant O-glycosylation.. http://www.ncbi.nlm.nih.gov/pubmed/9309422 ...
Protein Microarrays Based on Single Framework Recombinant Antibody Fragments (SINFABS) - Catcher and Carrier - A Crucial...
phdthesis{34b38b30-7926-4d3b-be18-4e6e97c97e27, abstract = {Antibody-based microarrays are among the novel class of rapidly evolving proteomic technologies. In recent years, antibody microarrays have emerged as a unique tool for high-throughput protein expression profiling with great promise within biomedicine and a wide range of potential applications, including disease diagnostics and biomarker discovery. In order to evolve the technology from small dedicated microarrays to high-density well-performing arrays for true global proteome analysis, this thesis focussed on the design of the two key components of the antibody microarray setup: the probe and the solid support, i.e. ?catcher and carrier?. The thesis is based upon five original papers that deal with i) the central features of the probe (on-chip functionality and stability, sensitivity and immobilisation strategies) and ii) the biocompatibility of the solid support (defined by the spot morphology, binding capacity, signal to noise ratio, ...
Antibodies | Free Full-Text | Dual Targeting of Tumor Cells with Bispecific Single-Chain Fv-Immunoliposomes | Notes
Antibody fragments, especially single-chain Fv fragments, have been established for the generation of immunoliposomes for targeted drug delivery in cancer therapy and other applications. Bispecific immunoliposomes should be useful for dual targeting addressing inter- and intratumoral heterogeneity of tumor antigen expression. Here, we established a protocol to generate dual-targeted immunoliposomes using genetically engineered scFv molecules recognizing two different tumor-associated antigens, EGFR and CEA (CEACAM5), applying a step-wise insertion of antibody-coupled micelles into preformed PEGylated liposomes. The dual-targeted immunoliposomes retained binding activity for both antigens and combined the selectivity of both antibodies within one liposome. Thus, these dual-targeted immunoliposomes should be suitable to deliver therapeutic payloads to tumor cells expressing EGFR or CEA, or both antigens.
Aminosyn II 8.5 % with Electrolytes, Sulfite-Free - Side Effects, Uses, Dosage, Overdose, Pregnancy, Alcohol | RxWiki
Aminosyn II 8.5 % with Electrolytes, Sulfite-Free - Get up-to-date information on Aminosyn II 8.5 % with Electrolytes, Sulfite-Free side effects, uses, dosage, overdose, pregnancy, alcohol and more. Learn more about Aminosyn II 8.5 % with Electrolytes, Sulfite-Free
Aminosyn II 4.25% in 10% Dextrose - Side Effects, Uses, Dosage, Overdose, Pregnancy, Alcohol | RxWiki
Aminosyn II 4.25% in 10% Dextrose - Get up-to-date information on Aminosyn II 4.25% in 10% Dextrose side effects, uses, dosage, overdose, pregnancy, alcohol and more. Learn more about Aminosyn II 4.25% in 10% Dextrose
Properties of a single-chain antibody containing different linker peptides<...
TY - JOUR. T1 - Properties of a single-chain antibody containing different linker peptides. AU - Alfthan, Kaija. AU - Takkinen, Kristiina. AU - Sizmann, Dorothea. AU - Söderlund, Hans. AU - Teeri, Tuula. N1 - Project code: BEL2026. PY - 1995. Y1 - 1995. N2 - Single-chain antibodies were constructed using six different linker peptides to join the VH and VL domains of an anti-2-phenyloxazolone (Ox) antibody. Four of the linker peptides originated from the interdomain linker region of the fungal cellulase CBHI and consisted of 28, 11, six and two amino acid residues. The two other linker peptides used were the (GGGGS)3 linker with 15 amino acid residues and a modified IgG2b hinge peptide with 22 residues. Proteolytic stability and Ox binding properties of the six different scFv derivatives produced in Escherichia coli were investigated and compared with those of the corresponding Fv fragment containing no joining peptide between the V domains. The hapten binding properties of different antibody ...
Patent WO2011044186A1 - Human single-chain t cell receptors - Google Patents
A soluble human single-chain T cell receptor (TCR) having the structure: Vα2-L-Vβ or Vβ-L-Vα2, wherein L is a linker peptide that links Vβ with Vα, Vβ is a TCR variable β region, and Vα2 is a TCR variable α region of the family 2 is provided. The provided scTCR is useful for many purposes, including the treatment of cancer, viral diseases and autoimmune diseases.
OPUS 4 | Search
Introduction: ScFv(FRP5)-ETA is a recombinant antibody toxin with binding specificity for ErbB2 (HER2). It consists of an N-terminal single-chain antibody fragment (scFv), genetically linked to truncated Pseudomonas exotoxin A (ETA). Potent antitumoral activity of scFv(FRP5)-ETA against ErbB2-overexpressing tumor cells was previously demonstrated in vitro and in animal models. Here we report the first systemic application of scFv(FRP5)-ETA in human cancer patients. Methods: We have performed a phase I dose-finding study, with the objective to assess the maximum tolerated dose and the dose-limiting toxicity of intravenously injected scFv(FRP5)-ETA. Eighteen patients suffering from ErbB2-expressing metastatic breast cancers, prostate cancers, head and neck cancer, non small cell lung cancer, or transitional cell carcinoma were treated. Dose levels of 2, 4, 10, 12.5, and 20 μg/kg scFv(FRP5)-ETA were administered as five daily infusions each for two consecutive weeks. Results: No hematologic, ...
OPUS 4 | Search
Introduction: ScFv(FRP5)-ETA is a recombinant antibody toxin with binding specificity for ErbB2 (HER2). It consists of an N-terminal single-chain antibody fragment (scFv), genetically linked to truncated Pseudomonas exotoxin A (ETA). Potent antitumoral activity of scFv(FRP5)-ETA against ErbB2-overexpressing tumor cells was previously demonstrated in vitro and in animal models. Here we report the first systemic application of scFv(FRP5)-ETA in human cancer patients. Methods: We have performed a phase I dose-finding study, with the objective to assess the maximum tolerated dose and the dose-limiting toxicity of intravenously injected scFv(FRP5)-ETA. Eighteen patients suffering from ErbB2-expressing metastatic breast cancers, prostate cancers, head and neck cancer, non small cell lung cancer, or transitional cell carcinoma were treated. Dose levels of 2, 4, 10, 12.5, and 20 μg/kg scFv(FRP5)-ETA were administered as five daily infusions each for two consecutive weeks. Results: No hematologic, ...
CiNii Articles - Cross-Neutralization Activity of Single-Chain Variable Fragment (scFv) Derived from Anti-V3...
The V3 loop in the envelope (Env) of HIV-1 is one of the major targets of neutralizing antibodies. However, this antigen is hidden inside the Env trimer in most isolates and is fully exposed only during CD4-gp120 interaction. Thus, primary HIV-1 isolates are relatively resistant to anti-V3 antibodies because IgG is too large to access the V3 loop. To overcome this obstacle, we constructed single-chain variable fragments (scFvs) from anti-V3 monoclonal antibodies 0.5γ, 5G2, and 16G6. Enhanced neutralization by 0.5γ and 5G2 scFvs was observed in strains resistant to their IgG counterparts. Neutralization coverage by 0.5γ scFv reached up to 90% of the tested viruses (tier 2 and 3 classes). The temperature-regulated neutralization assay revealed that extensive cross-neutralization of 0.5γ scFv can be explained by post-attachment neutralization. Neutralization assay involving viruses carrying an inter-subunit disulfide bond (SOS virus) showed that the neutralization-susceptible timeframe after ...
Conformation-Specific Blockade of the Integrin GPIIb/IIIa | Circulation Research
The adhesion-molecule group of integrins share the property to change conformation, on cell activation, thereby exposing their receptor ligand-binding pocket.2 This provides the unique possibility to design agents that specifically block only the activated receptor. We designed a strategy to develop human single-chain antibodies specific for integrins in the activated conformation that could be used for the therapy of the many diseases in which integrins play a major role, such as inflammation, cancer, and thrombosis.18 The platelet integrin GPIIb/IIIa is of pivotal importance for coagulation and thrombosis. Its conformation-unspecific blockade has been one of the major advances in antithrombotic therapy of recent years, although unexpected limitations and side effects, in particular with orally applicable blockers, have damped the original enthusiasm. We have taken advantage of the existence of different conformational states of GPIIb/IIIa to develop an alternative, unique strategy that targets ...
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Recombinant Antibody (scFv-Fc) Production Services - Syd Labs
Recombinant antibody (scFv-Fc) production services include: molecular construction of recombinant scFv-Fc antibodies; transient production of recombinant antibodies; purification of recombinant antibodies.
Characterization of a Single-Chain Variable Fragment Recognizing a Linear Epitope of Aβ: A Biotechnical Tool for Studies on...
JPT Peptide Technologies is a DIN ISO 9001:2015 certified and GCLP compliant integrated provider of innovative peptide based catalog products and custom services.
FCCT Biochemistry Lab:Research - OpenWetWare
Plasmid engineering (Marko Dolinar, Nejc Jelen) Plasmids are the prime gene vehicles in bacterial biotechnology. Understanding their mode of action is essential for designing improved vectors. We have successfully designed and built a synthetic biology based expression vectors we further used for production of single-chain antibody fragments in E. coli. Recently, we have constructed a hybrid shuttle vector that combines sequences of a cyanobacterial cryptic plasmid and of a synthetic biology vector. This is how we plunged into the sofisticated world of cryptic plasmids that cyanobacteria collected in course of evolution. (read more...) ...
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IRJPAS
Effective neutralization of different strains of HIV virus is shown by neutralizing antibodies against HIV-1 envelope glycoproteins. It has been demonstrated that gp41 is involved in virus mediated membrane fusion which results in HIV-entry into target cells. Recombinant single chain antibodies (scFvs) with high specificity and high affinity properties have been identified as useful agents in anti-viral targeted therapy. In this study we selected specific scFvs against a conserved neutralizing epitope of gp41. Four rounds of panning were performed to select the specific clones. The reactivity of the selected scFvs against the corresponding epitope was tested in ELISA. Results demonstrated that the specific clones were selected with the frequencies of 65% and 30%. The ELISA evaluation demonstrated significant higher OD of scFvs in reaction with the corresponding epitope than the negative control. Specific scFvs against conserved neutralizing epitope of gp41 of HIV has the potential to be ...
Multivalent and bispecific antibody fragments from E.coli - Research Collection
PROTEINFRAGMENTE (BIOTECHNOLOGIE); ANTIKÖRPER + IMMUNOGLOBULINE + GAMMAGLOBULINE (IMMUNOLOGIE); ESCHERICHIA (MIKROBIOLOGIE); PROTEIN FRAGMENTS (BIOTECHNOLOGY); ANTIBODIES + IMMUNOGLOBULINS + GAMMA GLOBULINS (IMMUNOLOGY); ESCHERICHIA (MICROBIOLOGY ...