CAJ | 학술논문

目的研制用于疟疾简便、快速诊断试剂盒的试剂.方法利用噬菌体抗体库技术,构建抗恶性疟原虫丝氨酸重复抗原(SERA)的噬菌体抗体库.经3轮"吸附-洗脱-扩增"的富集反应后,从中筛选抗SERA的阳性克隆株,并进行可溶性诱导表达,最后用ELISA和Western blot等进行鉴定.结果成功地构建了中等库容的噬菌体抗体库,并从中筛选到9株抗SERA的阳性克隆株,表达的单链抗体的相对分子质量大小约为31kDa,能与SE47′抗原起特异性结合反应.结论成功地构建了抗SE47′的噬菌体抗体库,从中筛选制备的单链抗体为研制疟疾快速诊断试剂盒奠定了基础.
목적연제용우학질간편、쾌속진단시제합적시제.방법이용서균체항체고기술,구건항악성학원충사안산중복항원(SERA)적서균체항체고.경3륜"흡부-세탈-확증"적부집반응후,종중사선항SERA적양성극륭주,병진행가용성유도표체,최후용ELISA화Western blot등진행감정.결과성공지구건료중등고용적서균체항체고,병종중사선도9주항SERA적양성극륭주,표체적단련항체적상대분자질량대소약위31kDa,능여SE47′항원기특이성결합반응.결론성공지구건료항SE47′적서균체항체고,종중사선제비적단련항체위연제학질쾌속진단시제합전정료기출.
Aim To develop new reagent for simple and fast diagnosis of plasmodium falciparum (p.f). Methods　　Phage-isplay antibody library technique was used to construct an antibody library, which is anti recombinant N-terminal main (SE47') of serine repeat antigen of p.f. Anti-SE47' phages were screened and enriched by panning with SE47' for three rounds of "adsorbing-diluting-proliferating". The positive phages were chosen to be induced for xpression soluble ScFvs. The antibodies' characteristic and specificity were determined by ELISA and Western blot. Esults　　The phage-displayed antibody library has been constructed and nine clones with good reactivity with SE47 in ELISA were selected. Soluble ScFvs were induced for expression and the results of Western blot showed that all could bind to SE47’ specifically and had a molecular weight about 31kDa. Conclusion　　The phage-displayed antibody library has been successfully constructed and the interested antibodies developed from it could be potentially useful for further research.