Artificially Synthesized Salidroside Antigen
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摘要: 合成红景天苷人工抗原，为进一步建立免疫学分析方法提供依据，采用高碘酸钠氧化法分别将红景天苷（Salidroside）、牛血清蛋白（BSA）与卵白蛋白（OVA）人工偶联成免疫抗原（SAL-BSA）和包被抗原（SAL-OVA），并利用紫外光谱法、SDS-PAGE法鉴定。用制备的人工免疫抗原免疫小鼠，用间接酶联免疫法测定抗体效价。结果成功制备了SAL-BSA、SAL-OVA两种抗原，SAL-BSA偶联比可达到7：1，在免疫后的小鼠血清中检出了抗红景天苷的抗体，效价为1：4 000。成功合成红景天苷人工抗原，为下一步单克隆抗体的制备及建立相应的免疫学分析方法奠定了基础。Abstract: To establish an immunoassay method on salidroside (SAL), it is necessary to firstly create a complete SAL antigen. This study applied the sodium periodate oxidation method to synthesize the immunizing antigen, SAL-BSA, and the coating antigen, SAL-OVA, which were positively identified by means of ultraviolet spectrum and SDS-PAGE. Subsequently, lab mice were immunized with the SAL-BSA to produce antiserum against SAL. The antibody titer was, then, determined by indirect ELISA. The results showed that SAL was conjugated with BSA/OVA in a ratio of 7:1, and the serum antigen titer reached 1:4 000. With the complete SAL antigen successfully synthesized, the preparation of monoclonal antibodies for the establishment of an applicable immunoassay methodology became possible.
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Key words:
- salidroside /
- artificial antigen /
- antibody
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图 1 SAL、BSA和SAL-BSA的紫外图谱(200～400 nm)
Figure 1. UV spectra of SAL, BSA and SAL-BSA (200 nm to 400 nm)
图 2 SAL、OVA和SAL-OVA的紫外图谱(200～400 nm)
Figure 2. UV spectra of SAL, OVA and SAL-OVA (200 nm to 400 nm)
图 3 完全抗原SAL-BSA的SDS-PAGE鉴定结果
注：M为蛋白marker; 1为BSA(100μg·mL-1)，2为SAL-BSA(100μg·mL-1)。
Figure 3. Identification of synthesized antigen SAL-BSA by SDS-PAGE
表 1 间接ELISA法测定红景天苷抗体的效价
Table 1. Titers of salidroside antibodies determined by ELISA
血清稀释度 阳性血清(SAL-BSA) 阴性血清 1:1000 3.8847 0.1305 1:2000 3.2953 0.1254 1:4000 2.4541 0.1219 1:8000 1.7154 0.0934 1:16000 0.6485 0.0959 1:32000 0.3420 0.0918 1:64000 0.1911 0.1034 1:128000 0.1380 0.01535 
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