Did I screw up my antigen with glutaraldehyde?

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Did I screw up my antigen with glutaraldehyde?

Postby BuckRimfire » Apr 25 2017 6:22 pm

I have a protein (~80 KD with some coiled-coil) that we tried previously to use as an immunogen with poor response. We are going to try a new rabbit, and want to modify/crosslink the protein to spice it up. To a couple mg of protein in 5 ml of buffer I added glutaraldehyde to 50 mM and let it react 75 min at room temperature. I may have seen a slight increase in turbidity, as I expected.

I thought I should quench the excess glutaraldehyde, so I then added glycine to 50 mM. Within a couple of minutes the solution developed a golden yellow color, which I guess is a polymer of glycine and glutaraldehyde. I dialysed the sample against PBS + 25% PEG to concentrate it, and the color became more concentrated, so whatever it is, it's too big to go through a 12,000 MWCO membrane.

Anyone know if this is going to affect the use in immunization?
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