Monoclonal antibody production
Review monoclonal antibody production technologies
Monoclonal antibody production can be performed by different technologies including monoclonal antibody production by hybridoma technology and by phage display technology. In this website we will explain how monoclonal antibodies can be generated by the hybridoma technology. Monoclonal antibody production by phage display technology is reviewed in our website www.phagedisplay.net
Review of monoclonal antibody production by the hybridoma technology
Monoclonal antibody production by the hybridoma technique consists in extracting B lymphocytes from the spleen of a laboratory animal (e.g. mice) that had been first exposed to an antigen to which we are interested in isolating an antibody against. The B cells can secrete monoclonal antibodies but not in a significant quantity and do not survive for long. The specific antibody producing B-cells are fused with myeloma cells which is a type of cancer cell line, which grow indefinitely in cell culture. Also the key in monoclonal antibody production by the hybridoma technology is the fusion of the two cell types, one is programmed to produce specific monoclonal antibodies but without the capacity of multiplying indefinitely and the other immortal with high capacity to grow but without the capacity of producing antibodies. The resulting cells, called hybridoma, will have the capacity to multiply quickly and indefinitely and to produce high quantity of monoclonal antibodies. During the process the cells are diluted to obtain one cell per well. The monoclonal antibodies produced by different clones are analyzed to determine their capacity to bind the antigen. By this way the positive clones are selected and isolated. See a picture of the hybridoma technology. The monoclonal antibody production can be done in cell culture or in laboratory animals. When the hybridoma cells are injected in the peritoneal cavity of a mouse, it will produce a tumor which synthesize a fluid rich in monoclonal antibodies called ascitic liquid.
Custom monoclonal antibody production services
VRISKO LTD delivers antibody services of high quality and to a low cost. We are committed to provide the highest level of products and custom monoclonal antibody production services.
We are a monoclonal antibody production company that is specialized in custom monoclonal antibody production services intended for recognition of target antigen in different forms. Monoclonal antibodies can be raised using various immunogens, such as peptides and recombinant proteins, however, in the end it is important that the obtained antibodies can recognize the target antigens in its native form e.g. protein antigens in tissue lysates. Vrisko Ltd has established a standard operating procedure in selectively identifying antibodies that bind to not only the immunogens but also the target antigens in ELISA, Western blotting, Immuno-precipitation, Immuno-fluorescence and Immunohistochemistry. Our strategy relies on our unsurpassed expertise in different fields and we have well-tested protocols in immunogen preparation, animal immunization and hybridoma screening
Our monoclonal antibody production services
The most important steps for successful monoclonal antibody production include antigen preparation, immunization approach, and screening strategy. Vrisko Ltd take great care of antigen designing and production, novel immunization strategy, and a careful mouse strain choice . Our monoclonal antibody production is divided into different steps, with a non-refundable part to cover the material cost to start the project. The balance will be due upon service completion.
- Immunisation and boosts for 5 Balb/c mice with standard immunisation protocol.
- Titer assay (ELISA) prior to selection of donor mice.
- Hybridoma fusion using splenocytes or lymph node cells and SP2/0 myeloma cells.
- Screening of up to 2,000 hybridoma clones for positives with ELISA. Up to 10 highly positive clones will be subcloned, expanded, and 5 ml of hybridoma culture medium will be provided to customer for evaluation.
- Customer selects 3 clones that will be expanded for cryopreservation and sent to the customer.
Antibody library against epitopes of a protein
A library of monoclonal antibodies against an array of antigens representative of a target protein can be generated, example the exposed epitopes of a surface protein.
A computational model is used to identify those residueues that are exposed on the protein surface by combining secondary structure prediction, phylogenetic analysis and structural modeling.
The identified epitopes are expressed as recombinant soluble proteins in E.coli.
Mice immunized with structural immunogens will produce antibodies that may recognize a few dominant conformational epitopes of the target proteins.
Contact us for more information of our custom monoclonal antibody production services.