I am quantifying the molar ratio of hapten to cbsa using maldi tof ms for my conjugation work. unfortunately,cbsa doesnt give any signal using maldi tof ms.but,tnbs shows the positive result (conjugation is happen).does anyone knows how to determine the hapten density (molar ratio hapten to protein) other than maldi tof ms.?
Dr,
I used sinapinic acid solution and alpha cyano matrix. Both unable to be detected
Yup.I did.but its not working.I need to find other method to support the result other than tnbs method.if anyone has suggestion,kindly plz make a comment :)
I would also test dihydroxibenzoic acid (DHB) or superDHB as matrices for BSA analysis.
Have you checked the following points?
- signal for your matrices alone;
- signal for your protein alone, I mean, not conjugated;
- concentration of your protein after conjugation;
- possible interfering reagents used for conjugation process that may affect MALDI;
- used zip-tip (C18) for clean-up.
These are some observations that we take in account to peptide analysis by MALDI.
If all of these are fine then you should consider other method.
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