anyone know something about making monoclonal antibodies?

What would be an expected final concentration of monoclonal antibody product in a batch. I’ve been told that 5-10 grams per liter is a fairly high concentration, but I want to see if I can confirm that.

So if I were in a company making monoclonal antibodies on a fairly large scale, what concentration range would I expect to see at the end of production?

Also, what about proteins in general? What concentration range would I see for a mammalian cell culture? (I know it “depends on the system”, I’m looking for a reasonable range or ranges)

Well I would have thought more science geeks would have been here

Can it be? A science question on the Dope with no answer?!

Well, I’ve done very little work with antibodies and nothing at all with mammalian cell culture, but I have done a bit with proteins in bacteria. From what little I remember, the ranges of concentrations for lab-purified protein were usually between a few mg/L to a few g/L, with most somewhere in between.

After looking around, it looks like you can buy commercial monoclonal antibodies at around 1-2 g/L. At least for the standard control antibodies we use – for the rest, we rarely use monoclonal.

eta: that concentration was for monoclonal anti-tubulin from Sigma.

By “batch”, are you referring to cell culture or mouse ascites? If the latter, you can produce quite a bit (though ascites production is not something I take lightly. It’s pretty awful for the mice, so I do it only if there is no other option.) Probably 20 mg, even as much as 50 mg per mouse.

If you’re doing it in cell culture, I would highly suggest roller bottles. 5-10 g/L sounds high even for a very experienced lab. I would say, if you do this all the time, expect 1-5 g/L. If you don’t, expect MUCH lower.

There’s reasons this sort of thing is generally outsourced to companies. I work for a biotech company and we do none of this in house. Much cheaper and easier to send off the hybridoma and get back the purified antibody.

What do you mean by “at the end of production”? It sounds like you may be more interested in the yield rather than the concentration as such?

In theory, it’s possible to concentrate protein(s) to a more-or-less arbitrary degree, dependent mainly on how much money you want to spend doing so and how much loss you are willing to tolerate during the process. The concentration of protein in monoclonal antibodies offered for commercial sale varies quite a lot, depending on the strength of the antibody and the intended use. As supplied to customers, they’re often lyophilized (dried down), which I guess is equivalent to a concentration of 100% (not really, since there’s usually some neutral carrier protein like BSA added and some salts/buffers too).

you have your vat (or roller bottle, or stainless steel tank of 10,000 liters), with a cell culture making these antibodies (or other protein…if antibody is inappropriate). After your cells are done with production, and BEFORE you start to purify the product from the media; what is the concentration of the protein/antibody in the cell culture media.

I’m aware you can purify the protein to essentially any concentration…I want to know what the typical highest concentration would be in the media, while the cells are producing it and before the protein/antibody gets purified.

If you’re skilled, and have a good producing hybridoma, your sups will contain 1-5 g/L. If you’re planning an experiment, I might plan to make twice as much sup as I would need.

(seriously though, contracting it out is cheaper and easier in the long run!)

I often purify proteins from bacteria like lazybratsche, if you want some comparison. The most that I’ve ever gotten of very pure protein (after nickel column, ion exchange, size exclusion) is somewhere around 300 milligrams per liter (it was a bacterial protein, but from a different species), and on average I see more like 5-10 mg/liter when working with human proteins expressed in bacteria (we usually grow up 6 to 12 liters). But again that is very pure sample of protein, such that it is just about the only protein you even even see on a gel. If you want dirty protein you could increase that a lot.

That’s part of the reason for this question. I’m going to be talking to the companies that do the actual work…the ones that get the contracts. I just want to fill in some of the HUUUUUGGGE gaps in my knowledge before I try to talk to them.