I am currently working on a project that requires the isolation of light chain from reduced IgG for bottam up proteomics-Mass spectrometry. Kindly provide insights or recommendations. Thanks!
Can't you use protein A/G beads to bind the heavy IgG chains and enrich them away? My remembering is that A/G doesn't bind the variable light chains
There are anti kappa and anti lambda chain antibodies on the market to isolate your target...
The light chains have a molecular mass of about 25 kDa and the heavy chains have a mass of about 50 kDa. That difference should make it possible to separate them under reducing conditions by gel filtration chromatography.
Here is a paper describing a method using ion exchange chromatography.
https://pubmed.ncbi.nlm.nih.gov/21183185/
As mentioned above by Dr. Albert Lee, you can use protein A or protein G to remove heavy chain. You can also use cation exchange chromatography to separated light chain from heavy chain, especially when the amount of the light chains you need is small since you can use an high resolution analytical cation exchange chromatography column.
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