氨苄青霉素人工抗原的制备及鉴定

Synthesis and Identification of Ampicillin Artificial Antigen

本研究采用戊二醛法和物理法制备氨苄青霉素(AMP)人工抗原,以紫外扫描和SDS-PAGE电泳鉴定偶联成功,采用改良的BCA法测定偶联率分别为8～10和17～21。通过单因素和L9(34)正交试验研究了反应物起始摩尔比、偶联剂的添加量和偶联时间等因素对偶联率的影响,选择最佳的偶联条件。最终确定戊二醛法的最佳反应条件为:反应物起始摩尔比40:1;戊二醛10μl;反应时间4h。物理法的最佳反应物起始摩尔比为20:1,反应时间3h。本研究表明物理法优于戊二醛法,是一种更好的制备氨苄青霉素人工抗原的方法。

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（3^4） 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 10μ1, 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.