23 Feb 2024
To test the affinity between protein A and multiple small molecules.

Technical difficulties:
The wild-type protein A tends to precipitate upon addition of high concentrations of reducing agents, rendering it unsuitable for SPR testing.
The protein gradually loses activity during the testing process.

By analyzing the protein sequence, truncating the protein, and introducing mutations to surface cysteine residues, a stable protein was obtained.
Initially, protein A was immobilized on the chip via amino coupling, but during the testing process, the protein gradually lost activity. Subsequently, protein capture was achieved using anti-His antibodies to immobilize the protein on the chip, ensuring its activity throughout the testing process.


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