Abstract: In this paper,ampicillin artificial antigens were synthesized by the methods of glutaraldehyde and physiological coupling,and were identified by UV scan and SDS-PAGE electrophoresis.The coupling rates are 8～10 and 17～21 respectively,both determined by the improved BCA method.Single factor test and orthogonal test design methods L9(34)were employed to study the influence of the factors such as the initial molar ratio of reagents,the addition of coupling agent and the coupling time to the coupling rate,as to determine the optimal reaction conditions for synthesis.The obtained results were that:the initial molar ratio of ampicillin to BSA 40:1,glutaraldehyde 10μl,coupling time 4 hours in glutaraldehyde method,in physical method 20:1 and coupling for 3 hours.Results indicated that the physiological method is superior to others for preparation of ampicillin artificial antigens.

Key words: ampicillin, artificial antigen, coupling rate