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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?

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  • $\begingroup$ If it is a monoclonal Ab you are after, I think you will need to get it commercially, either by buying the antibody or by supplying an antigen to a commercial company (too difficult and expensive to set up in lab). The other option is a polyclonal Ab, where you will need to inject pure Ag into (say) a rabbit and (at the very least) isolate the IgG fraction from serum (and beware of antibodies produced against contaminants present in the 'pure' antigen preparation) [You don't, of course, need pure Ag for monoclonal Ab production]. But what are you designing? An ELISA? $\endgroup$ – user1136 Mar 19 at 12:42
  • $\begingroup$ If it is not that difficult in your organism, then you can create a transgenic with your protein of interest with a HA or a FLAG tag. The anti-HA and anti-FLAG antibodies are cheap. $\endgroup$ – WYSIWYG Mar 22 at 12:33
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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/

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