Murine T cell hybridomas bearing receptors for antigen plus I region gene products were used as immunogens in mice in an effort to raise anti-receptor antisera. The antisera were assayed for anti-receptor activity by the ability to inhibit interleukin 2 production by the T cell hybridomas stimulated by antigen and I region expressing antigen-presenting cells. The T cell hybridomas used in these experiments were made by fusing antigen-specific, I region-restricted BALB/c T cell blasts to the AKR thymoma, BW5147. Three groups of mice were immunized with the T cell hybridomas: (BALB/c X AKR)F1 animals, syngeneic to the hybridoma; (BALB.B X aKR)F1 animals, differing from the hybridomas at H2; and (C.B20 X AKR)F1 animals, differing from the hybridomas at Igh. Mice were immunized multiple times and sera from individual animals were assayed for anti-receptor antibodies. In all groups, some mice produced anti-receptor antibodies by the criterion that they were inhibitory in the assay mentioned above. ...
TY - JOUR. T1 - Analysis of rheumatoid factors by a biotin-avidin based isotype-specific ELISA. AU - Kemp, Mette. AU - Husby, Sofie. AU - Jensenius, Jens Christian. AU - Rasmussen, G G. AU - Svehag, S E. PY - 1985. Y1 - 1985. N2 - A one day enzyme immunoassay for the detection of rheumatoid factors of the IgG, IgM, and IgA class is described. The assay utilizes rabbit IgG as solid-phase reactant and the biotin-avidin interaction for the coupling of enzyme to indicator antibody. Three different indicator antibodies discriminated effectively between rheumatoid arthritis patients and normal subjects. F(ab)2 fragments of goat antibodies were found best suited for the test. Rheumatoid factor activity was expressed in U/ml by comparing samples to an internal standard, which was related to the WHO international reference serum for rheumatoid arthritis. Rheumatoid factor activity (U/ml) in the IgM-specific assay showed a close correlation to the latex agglutination titer. Avidity indices estimated from ...
Hybridoma-derived idiotype vaccines have been used for the experimental treatment of human lymphoma over the last twenty years, providing evidence of biological efficacy, clinical efficacy and clinical benefit. However, the product that has come closer to regulatory approval is unlikely to clear that hurdle due to the insufficiently robust data obtained in a recently closed clinical trial. This review aims at discussing the reasons for hybridoma-derived idiotype vaccines, more difficult to produce but also more successful than recombinant idiotype vaccines so far, are unlikely to gain regulatory approval. In particular, it is necessary to examine the many peculiar features of this therapeutic approach in a broader context, with special attention to concepts like customized active immunotherapy and randomization. Most published trials based on hybridoma-derived idiotype vaccines are being analyzed, together with the yet non-peer reviewed data from the only randomized study conducted so far with this
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TY - JOUR. T1 - Analysis of T-cell hybridomas with an unusual MHC class II-dependent ligand specificity. AU - Mendiratta, S. K.. AU - Singh, Nagendra. AU - Bal, V.. AU - Rath, S.. PY - 1996/1/1. Y1 - 1996/1/1. N2 - We have characterized two unusual T-cell hybridomas, 1E3 and 3B8, from H-2(k) mice immunized with I-Ab-transfected L cells (H-2(k)), that are stimulated by L cells transfected with I-Ab, I-A(k) or I-Eb, but not by non-transfected L cells. These hybridomas could not be stimulated by spleen cells from H-2(i3), H-2(k), H-2b or H-2(d) mice. Monoclonal anti-I-A antibodies did not block their responses, suggesting that mouse major histocompatibility complex (MHC) class II molecules may be peptide donors rather than restriction elements for them. The stimulation of these hybridomas by fibroblast targets was not blocked by an anti-H-2k(k),D(k)-specific monoclonal antibody. Lipopolysaccharide (LPS)-activated splenic and peritoneal exudate cells from H-2(k), H-2(d), H-2(i3), H-2b as well as ...
TY - JOUR. T1 - Changes of monosaccharide availability of human hybridoma lead to alteration of biological properties of human monoclonal antibody. AU - Tachibana, Hirofumi. AU - Taniguchi, Kiyotaka. AU - Ushio, Yoshitaka. AU - Teruya, Kiichiro. AU - Osada, Kazuhiro. AU - Murakami, Hiroki. PY - 1994/1/1. Y1 - 1994/1/1. N2 - The effect of glucose and other monosaccharide availability in culture medium on production of antibody by human hybridomas has been studied. Human hybridoma cells C5TN produce an anti lung cancer human monoclonal antibody, and the light chain is N-glycosylated at the variable region. When the cell line was grown in the presence of various concentrations of glucose, the antibodies produced changed their antigen-binding activities. Analysis of the light chains produced under these condition revealed that four molecular-mass variant light chains ranging from about 26 to 32 kDa were secreted. The twenty six-kDa species, which corresponds to a non-glycosylated form of the light ...
We have examined the ability of hCD4 to interact functionally with mouse class II MHC molecules using the mouse T cell hybridoma BI-141, specific for beef insulin. We have previously shown that expression of mouse CD4 results in a marked enhancement of IL-2 release by BI-141 cells in response to beef insulin or, in a cross-reactive response, to pork insulin, on the appropriate mouse APCs. We now demonstrate that expression of hCD4 results in an equivalent stimulation of antigen responses by this mouse T cell hybridoma. The specificity of this effect was demonstrated by mAb and gp120 blocking studies. These data provide the first direct evidence for function of hCD4 and in an exclusively mouse system. ...
A murine hybridoma cell line has been established that consistently forms large cytoplasmic inclusions. These structures bind antibody specific for mouse kappa L chain when stained in situ. SDS-PAGE analysis of isolated inclusion bodies produce a single protein band of approximately 26,000 Mr that reacts with anti-kappa antibody when transferred to nitrocellulose. No carbohydrate was detected in association with the purified protein. These data are consistent with the intracellular retention and deposition of complete kappa L chain protein. ...
TY - JOUR. T1 - Detection and characterization of murine ecotropic recombinant virus in myeloma and hybridoma cells. AU - Deo, Y.. AU - Ghebremariam, H.. AU - Cloyd, M.. PY - 1994. Y1 - 1994. N2 - Ecotropic recombinant virus (ERV), a relatively new class of murine retrovirus endogenous to mice, is expressed at significant levels by most murine myeloma and hybridoma cells examined. The routine XC, S+L-, mink cell focus-inducing (MCF), and reverse transcriptase (RT) tests are not suitable to detect and quantify the levels of ERV. A serological focus assay, based on specific anti-murine leukemia virus (MuLV) viral envelope (env) antibodies, is required to detect ERV. A more sensitive format of this serological focus assay includes co-cultivation of test article cells with the indicator (Mus dunni) cells. ERV isolated from murine hybridoma cells show a unique pattern of cross-reactivity with anti-MuLV env antibodies and this pattern is clearly distinct from that of ectropic and xenotropic ...
Kanagawa, O.; Nakauchi, H.; Sekaly, R.P.; Maeda, K.; Takagaki, Y., 1990: Expression and function of the transfected CD8 alpha chain in murine T cell hybridomas
MPs Opti-Clone™ is a partially purified hybridoma growth medium supplement which improves the cloning efficiency of murine B-cell hybridomas. It will enhance the growth of hybridomas cultured at low cell densities, and it will dramatically increase the number of antibody producing colonies during HAT selection. It supports the growth of hybridomas used in the manufacture of monoclonal antibodies. The formulation is suitable for use in cloning and fusion applications. Features: ∙ Promotes hybridoma growth ∙ Eliminates feeder cell layers ∙ Increases antibody production ∙ Improves stressed cells viability ∙ Improves surviving hybridomas ...
Expression of cellular oncogenes was studied in a T cell hybridoma that undergoes cytolytic activation when stimulated by specific antigen or by anti-Thy-1 antibody. The activation occurs without induction of hybridoma proliferation, providing a model to examine oncogene expression during functional differentiation of lymphocytes. We found that c-fos and c-ets-1 mRNAs were transiently induced at high levels in the hybridoma 30 min and 4 h after stimulation, respectively. c-myc and c-ets-2 oncogenes were constitutively expressed in the hybridoma and their mRNA levels were unaffected during 4 h of stimulation, although c-myc expression was reduced in the later stage of stimulation. Inhibitors of T cell activation, cyclosporin A and anti-LFA-1 antibody, blocked the induction of c-fos and c-ets-1 mRNAs without reducing the levels of c-myc and c-ets-2. The results indicate that the functional activation of the CTL hybridoma is associated with induction of c-fos and c-ets-1 genes. ...
The aim of this study was to establish hybridomas capable of long-term production of human monoclonal antibodies (mAbs). Heterohybridization was performed between the mouse myeloma cell line P3X63Ag8.653 and activated human peripheral blood lymphocytes (PBL). In order to achieve better retention of human chromosomes, as well as to improve the stability of the heterohybrids, one HAT-sensitive immunoglobulin (Ig)-non-secreting human x mouse (h x m) heteromyeloma was fused for a second time with activated human PBL. In this way, a panel of HAT-sensitive Ig-non-secreting h x h x m heteromyelomas was obtained and tested for its ability to generate stable human Ig-secreting heterohybrids with activated human PBL. Six lines were selected on the basis of their enhanced characteristics of fusion efficiency and genetic stability. When fused with in vitro immunized human PBL, they generated several h x h x h x m hybridomas stably secreting high yields (10-23 micrograms/ml/24 h) of human mAbs reactive with ...
TY - JOUR. T1 - Transmural pressure induces IL-6 secretion by intestinal epithelial cells. AU - Kishikawa, H.. AU - Miura, S.. AU - Yoshida, H.. AU - Hirokawa, M.. AU - Nakamizo, H.. AU - Higuchi, H.. AU - Adachi, M.. AU - Nakatsumi, R. C.. AU - Suzuki, H.. AU - Saito, H.. AU - Ishii, H.. PY - 2002. Y1 - 2002. N2 - Although intestinal epithelial cells (IECs) are known as an important source for IL-6, it is not known whether mechanical forces affect IL-6 production. We investigated how transmural pressure modulates IL-6 synthesis and activation of transcription factors in IECs. Pressure was loaded onto IEC-18 cells by introducing compressed helium gas into the cell culture flask for 1-48 h. IL-6 release into the culture media was determined by cell proliferation bioassay using an IL-6-dependent mouse hybridoma cell line (7TD1). Exposure to pressure (80mmHg) significantly enhanced IL-6 release into the culture media from IEC-18 cells at 12 h. Under control conditions, IL-6 secretion was directed ...
article{9a5c9918-f28d-42f7-9e4a-e617d47af020, abstract = {Development of type-II collagen (CII)-induced arthritis (CIA) is dependent on a T-cell mediated activation of autoreactive B cells. However, it is still unclear if B cells can present CII to T cells. To investigate the role of B cells as antigen-presenting cells (APCs) for CII, we purified B cells from lymph nodes of immunized and nonimmunized mice. These B cells were used as APC for antigen-specific T-cell hybridomas. B cells from naïve mice did present native, triple-helical, CII (nCII) but also ovalbumin (OVA) and denatured CII (dCII) to antigen-specific T-cell hybridomas. In addition, B cells primed with nCII or OVA, but not dCII, activated the antigen-specific T-cell hybridomas two to three times better than naïve B cells. We conclude that antigen-primed B cells have the capacity to process and present CII to primed T cells, and antigen-primed antigen-specific B cells are more efficient as APC than naïve B cells. We further ...
Read PDF to know how XP Media & CloneMedia for Mouse Hybridoma Generation provide a complete solution that supports all stages of hybridoma cell line development.
False-colour scanning electron micrograph of a hybridoma cell producing a monoclonal antibody to cytoskeleton protein. Monoclonal antibodies are produced by injecting a mouse with an antigen & harvesting its antibody response by removing the B lymphocytes. These are fused with myeloma (tumour) cells taken from a mutant lymphocyte source. The fusion product is called a hybridoma cell. B lymphocytes are short-lived; when fused with tumour cells, which divide indefinitely, the continuous production of antibody is secured. Hybridomas are screened for the required antibody; they are then cloned & produce that antibody. Magnification: x1500 at 35mm size. - Stock Image G400/0031
... are immortalized cells derived from the fusion of B lymphoblasts with a myeloma fusion partner. Some hybridomas in the ATCC collection are somatic cell hybrids. These cells are capable of producing immunoglobulins that are specific for viral, bacterial or cellular targets.
Adaltis can produce purified and not purified antibody by using its cell lines. Antibodies can be purified on request by: HPLC , FPLC , affinity chromatography and gel filtration.. For production service information please contact: [email protected] ...
The variable-region genes of monoclonal antibody against spores were cloned from mouse hybridoma cells by reverse transcription-PCR. food spoilage (9). Control of the bacterial spores in meals processing is vital that you ensure the basic safety and an extended shelf lifestyle of foods. To keep the product quality and basic safety of foods, polyclonal and […]. ...
There is disclosed a polypeptide (CD40-L) and DNA sequences, vectors and transformed host cells useful in providing CD40-L polypeptides. More particularly, this invention provides isolated human and murine CD40-L polypeptides that bind to the extracellular binding region of a CD40 receptor. Also disclosed are methods of simulating hybridoma cells to increase monoclonal antibody production by administering a CD40 ligand polypeptide that stimulates B cell proliferation.
METHODS OF PRODUCING HYBRIDOMAS AND MONOCLONAL ANTIBODIES AND ANTIBODIES PRODUCED THEREBY - diagram, schematic, and image 150 ...
The present invention pertains to the novel hybridoma SDW18.1.1, hybridomas obtained from SDW18.1.1, monoclonal antibodies obtained from such hybridomas and derivatives of such monoclonal antibodies. The novel hybridomas are formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with cachectin/TNF. Diagnostic and therapeutic utilities for the monoclonal antibodies and their derivatives are proposed, and testing procedures, materials in kit form and pharmaceutical compositions are likewise set forth.
Anti-ABeta Globulomer Antibodies, Antigen-Binding Moieties Thereof, Corresponding Hybridomas, Nucleic Acids, Vectors, Host Cells, Methods of Producing Said Antibodies, Compositions Comprising Said Antibodies, Uses Of Said Antibodies And Methods Of Using Said Antibodies - diagram, schematic, and image 78 ...
The DO11.10 mouse model is a valuable tool for studies of T cell immigration, immunoregulation, development, activation, and function. The KJ1-26 monoclonal antibody reacts with the T cell receptor (TCR) expressed on lymphocytes of the DO11.10 transgenic mouse and the TCR of the BALB/c-derived DO11.10 and DO11.10.24 T cell hybridoma. The DO11.10 TCR is specific for the chicken ovalbumin (OVA) peptide (323-339) in the context of I-A[d] major histocompatibility (MHC) molecules. Transgenic DO11.10 T cells also recognize OVA peptide from jungle fowl and turkey in the presence of A20-1.11, the H-2d-bearing, antigen-presenting B cell lymphoma. - Danmark
In a previous article, RAMPITSCH at BCRSSU.AGR.CA wrote: ,Hello nets: ,Im not sure if this is the right place to ask, but ..... ,I have a hybridoma cell line contaminated with fibroblasts. Normally I dont ,have a problem eliminating these since they stick to the plate a lot better ,than the hybridomas do: simply transferring the cells to a new plate usually ,does the trick. Now I have a line of fibroblasts which refuses to die (even ,after two months and more transfers than I care to remember). The hybridoma ,cell line (needless to say an important one) is growing poorly as it is barely ,able to attach to the plate and cant compete with these aggressive fibro- ,blasts (maybe they have been transformed?). Is there a miracle anti-fibro- ,blast agent? Is there any way of selectively getting rid of these fibroblasts? , ,Thanks for any help and comments, ,Chris R. , Chris: I may be brain-cramping but cant you just sort out the hybridomas? Or even pan them? Use something like an anti-Ig-FITC ...
Gibbons, J J.; Kim, Y T.; and Siskind, G W., "Regulation of hybridoma antibody production by coculture with antigen specific or idiotype specific immune spleen cell. Abstr." (1982). Subject Strain Bibliography 1982. 1805 ...
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Hybridoma cells usually grow to fairly low cell densities in batch cultures (1-3×106 cells/ml). The reason for this is either that essential nutritional components of the medium are consumed, or that
1. It is the disease caused by the novel coronavirus. Patients may have fever with respiratory symptoms - cough, runny nose, sore throat or difficulty breathing. It can even be fatal if the symptoms are severe.. 2. Route of Transmission: Human-Human transmission through droplets from coughing and sneezing of infected person and close contact with COVID-19 cases.. 3. Incubation period: 2-14 days. ...
An international team has developed an AI algorithm with social skills that has outperformed humans in the ability to cooperate with people and machines in playing a variety of two-player games. The researchers, led by Iyad Rahwan, PhD, an MIT Associate Professor of Media Arts and Sciences, tested humans and the algorithm, called S# ("S sharp"), in three types of interactions: machine-machine, human-machine, and human-human. In most instances, machines programmed with… read more. ...
An international team has developed an AI algorithm with social skills that has outperformed humans in the ability to cooperate with people and machines in playing a variety of two-player games. The researchers, led by Iyad Rahwan, PhD, an MIT Associate Professor of Media Arts and Sciences, tested humans and the algorithm, called S# ("S sharp"), in three types of interactions: machine-machine, human-machine, and human-human. In most instances, machines programmed with… read more. ...
The Gothenburg-based artist Nino Mick suggests in one of their poems that ultimately "biology is our only home". The film programme suggests different ways of approaching such an understanding of biology as home. This film programme consists of a screening of two movies: Pojktanten and a short film/installation, Sporing Lips of Transposed Desire. Each of these two audio-visual artefacts explore floral-aesthetics through something that can be rendered as plant-human intimacies. The programme is inviting the audience as entrants - more than simply viewers - to engage in these ecological intimacies and to think of the ethics and potentials of thinking intimacies as beyond a human-human relationality. Approaching the films through such an angle of ecological intimacy might allow to perceive a decentred understanding of the human through multiplicities and entanglements with others. Wibke will offer in a short introductory talk to the programme as a thinking tool to apply to the film ...
During the therapy session, visual 3D motion tracking will be used via Kinect, audio will be recorded, and basic measurements (i.e. the distance between a participants wrist and elbow) will be taken. After the patient-therapist interaction has been mapped, members of the research team will build computer based models of the therapist and patient using the insights gained from the human-human interaction. The models developed in this proposed study will be used as a template for programming safe and intuitive humanoid-patient interactions for future study. The model of the therapist will be implemented on the Robot (w/o a patient involved) and in a simulation environment where it will be tested with a computer-based model of the patient. ...
mouse anti-estrogen receptor alpha, ligand binding domain (aa 304-554) hybridoma (ERalpha BZ1) is an eagle-i resource of type Hybridoma cell line at eagle-i Network Shared Resource Repository.
mouse anti-late bloomer hybridoma (10C9 anti-late bloomer) is an eagle-i resource of type Hybridoma cell line at eagle-i Network Shared Resource Repository.
The T cell receptor (TCR) is a heterodimer composed of two transmembrane glycoprotein chains, α and β. Both chains are members of the Ig superfamily and consist of a constant and a polymorphic variable region. The variable region of the TCRα/β is involved in recognition of antigenic peptides presented by the MHC complex of antigen-presenting cells. The Anti-TCRα/β antibody recognizes a common determinant of the TCRα/β-CD3 complex. - Lëtzebuerg
After obtaining either a media sample of cultured hybridomas or a sample of ascites fluid, the desired antibodies must be extracted. Cell culture sample contaminants consist primarily of media components such as growth factors, hormones and transferrins. In contrast, the in vivo sample is likely to have host antibodies, proteases, nucleases, nucleic acids and viruses. In both cases, other secretions by the hybridomas such as cytokines may be present. There may also be bacterial contamination and, as a result, endotoxins that are secreted by the bacteria. Depending on the complexity of the media required in cell culture and thus the contaminants, one or the other method (in vivo or in vitro) may be preferable. The sample is first conditioned, or prepared for purification. Cells, cell debris, lipids, and clotted material are first removed, typically by centrifugation followed by filtration with a 0.45 µm filter. These large particles can cause a phenomenon called membrane fouling in later ...
Hybridoma technology is used to fuse fusion a B cell and myeloma to form a hybridoma that produces identical monoclonal antibodies.
Hybridoma technology is used to fuse fusion a B cell and myeloma to form a hybridoma that produces identical monoclonal antibodies.
Combining our 15 years of experience in gene synthesis, peptide synthesis, protein production and antibody development is the key to succeed in almost all projects. Indeed, we take into consideration parameters from these 4 services to imagine the best development strategies.. We all know how important it is to be able to produce the most appropriate antigen to succeed in developing the best hybridomas. But it is also important to select the best clones in the targeted application. Our guarantee that the clones will work in your experiment (your application and samples) ensures you that well use all means and efforts to deliver hybridomas producing the perfect antibodies you need.. Take part of this revolution by clicking the box below and we will study your project and send you an offer. ...
Hybridoma fusion and selection reagents are designed to assist fusion and select hybridomas from among normal cell populations, respectively.
I will soon be preparing a hybridoma with rat spleenocytes, and want a protocol for counting the spleenocytes, and how many spleenocytes to how many myeloma cells ...
The diversity of ATCC products includes the microbiology collection of organisms from bacteria, fungi, yeast, protozoa, animal viruses; molecular genomics materials (DNA/RNA, clones, vectors) and our cell biology products for primary cells, stem cells, tissue biology, and cell lines and hybridomas are included in our wide range of products used by researchers and industry in a variety of applications (assay testing of antibiotics, cell surface
TY - JOUR. T1 - Analysis of suppressor T cells induced by donor-specific transfusion (DST). T2 - establishment of a human T cell hybridoma producing an antigen-nonspecific suppressor factor.. AU - Fujiwara, T.. AU - Sakagami, K.. AU - Kusaka, S.. AU - Uda, M.. AU - Orita, K.. PY - 1992. Y1 - 1992. N2 - Formation of suppressor T cells (Ts) induced by donor-specific transfusion (DST) is one of the most commonly suggested mechanisms for the beneficial effect of DST. In this study, we established a human T cell hybridoma derived from the peripheral blood lymphocytes (PBL) of a DST-treated patient, which produced an antigen-nonspecific suppressor factor. Post-DST PBL were fused with an azaguanine-resistant mutant of a human T cell leukemia cell line, CCRF-CEM(AG). After selection and cloning, we established one clone producing the mixed lymphocyte reaction (MLR) inhibitory factor (C524: 18%-43% suppression). Suppressive activity of the supernatant obtained from C524 after activation by PHA was highly ...
Cesar Milstein and Georges J.F. Kohler invented the production of monoclonal antibodies in 1975 for which they got the Noble Prize of 1984 for Medicine and Physiology along with Niels Kaj Jerne, who made other contributions to immunology. The term hybridoma was introduced by Leonard Herzenberg in 1976-1977. Principle: The method starts by injecting a mice with immunogen. The mice should response to immunogenic reaction. A kind of somatic cell, the B-cell, produces antibodies that bind to the immunogen. These recently created antibodies are then collected from the mice. These isolated cells are merged with immortal B-cells (a myeloma cell), in order to provide a hybrid cell known as Hybridoma. These hybridomas have the antibody producing ability of the B-cell and also have the reproducibility and longevity. The hybridomas are fully grown in culture medium. In addition, the manufactured antibodies are all chemically identical in distinct to polyclonal antibody.. Hybridoma Production of Monoclonal ...
The most comprehensive custom mouse monoclonal antibody production services: custom production of monoclonal antibodies against non-modified, phospho-specific, methylation-specific, and acetylation-specific peptides and proteins; monoclonal antibodies for ELISA, Western blot, IP, IF, ICC, and IHC applications. Guaranteed ELISA > 1:40 000.
TY - JOUR. T1 - Monoclonal antibodies to various morphologic components of human skin. AU - Aiba, S.. AU - Masuko, T.. AU - Hosokawa, M.. AU - Hashimoto, Y.. PY - 1983/1/1. Y1 - 1983/1/1. N2 - Somatic cell hybrids were established from the mouse myeloma, P3x63Ag8.653 cells, and the spleen cells of a mouse hyperimmune to human epidermal cells. Indirect immunofluorescence test with hybridoma culture fluids displayed that 253 out of 263 hybridoma cultures secreted antibodies reactive with the frozen sections of human skin. The hybridomas secreting unique antibodies to skin components were cloned and designated as AHS-1 to -8. Monoclonal antibodies (MoAb) from AHS-1, AHS-2, and AHS-3 hybridomas did detect cytoplasmic antigens present in the epidermal layer, eccrine ducts and glands (except MoAb AHS-1), and hair follicles. Enzyme-linked immunosorbent assay showed that the antigen recognized by either MoAb AHS-1 or MoAb AHS-2, but not by MoAb AHS-3, shares the antigenic determinant with antigen(s) ...
Monoclonal antibodies (MAbs) are made by identical immune cells and target one particular epitope by monovalent or monospecific affinity. The high affinity and selective binding of MAbs to epitopes in target antigens makes them highly potent tools for use in biochemistry, molecular biology and medicine. The first working method described for the isolation of monoclonal antibodies was hybridoma technology, based on forming hybrid cell lines (hybridomas) by fusing an antibody-producing B-cell with a myeloma cell [1]. The antibodies produced by a particular hybridoma clone share the same specificity. Thus, individual clones can be screened for the production of an antibody with the desired affinity. However, hybridoma technology has shortcomings: it takes a relatively long time (on the order of months) and has not been widely applied to organisms other than mice. Moreover, antibody sequence information is unavailable by this method. Thus, when a hybridoma-screened antibody is selected for further ...
Our hybridoma bank contains over 20 murine hybridomas. Moreover, we have extensive experience in the purification and characterization of numerous murine monoclonal antibodies for in vitro and in vivo use. We have also generated monoclonal antibodies de novo by in vivo immunization and subsequent in vitro fusion of heterologous fusion partners from Chinese hamsters and rats, and produced IgA-secreting hybridomas de novo. Depending on the quantity of monoclonal antibodies required, hybridomas are propagated in either static flat tissue culture flasks or in roller bottle cultures. Monoclonal antibodies are purified by affinity column chromatography using protein G charged columns. The affinity purified monoclonal antibodies are dialyzed against deionized water, concentrated on a vacuum concentrator, resuspended in sterile PBS, and the protein concentration determined by BCA. The purity of the monoclonal antibodies is confirmed by western blotting with commercial anti-mouse IgG or IgM, antibodies, ...