Analysis of Monoclonal Antibody

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Dr. Analytical
Dr. Analytical's picture
Analysis of Monoclonal Antibody

Greetings from the Analytical Chemistry forum!

I need a little (OK, maybe a lot) general information on monoclonal antibodies. Being a chemist with only some biochemistry in my background, I have been asked to assist with an analysis of a monoclonal antibody product. I believe one of the questions relates to aggregation, and the lab will be using a chromatographic technique to assess this (that's where I come in). At this time I do not have any specifics on the material; that will come later.

What I need now is a little help getting started. Is there a primer or general introduction to this field that I can get somewhere? I would like information on how these things are made, their structure, and behavior in solution. I realize there are thousands (millions?) of possible materials, so at this point I would like to get a sense on what the field is about in its most general terms, and get an idea on the range of antibodies that are out there. From there, I will be able to see where this particular one fits into the grander scheme.

Thanks in advance!

Dave@AbDSerotec
Dave@AbDSerotec's picture
I hate to suggest it on a

I hate to suggest it on a scientific forum, but the wikipedia entry on 'Antibody' (http://en.wikipedia.org/wiki/Antibody)actually is a nice primer. Lots of links to references and external sites as well. Also check out the wiki entries on 'Monoclonal Antibody' and 'Hybridoma.'

Feel free to PM or email me with any specific questions and I'll try my best to help you out.

Good luck!

Jose Rodríguez
Jose Rodríguez's picture
So I need a similar help with

So I need a similar help with this. I fused my hybridoma cells and I was able to see positive labeling in tissue with those Monoclonal antibodies. Now characterize those monoclonal antibodies some of the techniques used are western blots and chromatography, but what other techniques can I use to characterize it and have more information abour it?.

thanks for the help.

samm
samm's picture
ELISA - subtyping; rtPCR

ELISA - subtyping; rtPCR analysis - plus the chromatographic purification, Western and IHC you've already done will be ample.

Sami Tuomivaara
Sami Tuomivaara's picture
Jose,

Jose,

ELISA (see for example) is the most widespread tool for antibody characterization. The ultimate target is to characterize the epitope(s) of the antibody, so you can structurally interpret immunolocalization and other experiments. ELISA is the way to go here, especially in the screening phase, because you can do it in high-throughput manner. The abomentioned wikipedia articles are helpful as well.
Cheers,