How are primary monoclonal antibodies for screening mutant cells made, physically?

Question

I'm working with a fairly common protein expressed in a large numbers of organisms, let's say a keratin-associated beta-protein. I'm trying to develop a process which requires primary-secondary antibody screening for the keratin-associated beta-protein. Trouble is, it's expensive. Very much so.

I can't find the antibody for any less than $778/mL, and after accounting for dilution and volume required for the recommended protocol, I can't imagine I'll be able to finish designing the process with any less than 20mL. The protein can't change as a design constraint, so there's no reason I'd ever need to change my antibody, but if I need so much... is there any way I can make it myself?

How are these antibodies actually manufactured? Would I need to get ahold of actual B cells and use those, or could I isolate the cellular machinery that produces the antibodies and use that?

Answer 1

Monoclonal antibodies are typically manufactured from a hybridoma cell line. If you are able to obtain hybridomas already made for your antibody (not very likely), or another antibody against your protein, then it's quite easy to produce the antibody yourself. The hybridomas secrete the antibodies into the media, so after growing a bunch of cells, you can purify the antibody from the media with a protein G column.

However, there are not very many hybridoma cell lines available. It's a lengthy and expensive process to make your own (several months and thousands of dollars), even with the help of a core facility/company. You would provide the facility with several milligrams of your protein, they would use it to immunize mice, then after 1-3 rounds of screening, you could obtain hybridoma clones.

There are some banks with hybridomas available for research, for example:

http://dshb.biology.uiowa.edu/

http://www.cellbankaustralia.com/ecacc-hybridoma/