中德临床肿瘤学杂志(英文版)
中德臨床腫瘤學雜誌(英文版)
중덕림상종류학잡지(영문판)
THE CHINESE-GERMAN JOURNAL OF CLINICAL ONCOLOGY
2003年
1期
39-41
,共3页
肖锡宾%张昌卿%张颖%张如华%李经略%冯凯涛%孙韵%叶永照
肖錫賓%張昌卿%張穎%張如華%李經略%馮凱濤%孫韻%葉永照
초석빈%장창경%장영%장여화%리경략%풍개도%손운%협영조
抗原表位%随机多肽文库%单克隆抗体%鼻咽癌
抗原錶位%隨機多肽文庫%單剋隆抗體%鼻嚥癌
항원표위%수궤다태문고%단극륭항체%비인암
epitope%random peptide library%monoclonal antibody%nasopharyngeal carcinoma
目的 鉴定能被BAC5单抗识别的定位于鼻咽癌细胞表面的抗原表位.方法 应用BAC5单抗作为靶抗体对噬菌体呈现的随机12肽文库进行3轮生物淘洗,用抗体捕获和竞争试验的夹心ELISA方法选择和鉴定阳性噬菌体克隆,对阳性和阴性噬菌体的外源性DNA片段进行序列分析,推导和比较由这些噬菌体所呈现的多肽氨基酸序列.结果 通过3轮生物淘洗能被抗体捕获的噬菌体克隆为77%(35/45).用竞争试验从所捕获的克隆中测得8个阳性克隆.来自这8个克隆的噬菌体呈现三种外源多肽,即-H-Q-S-H-Y-P-Y-P-V-V-S-L-(4/8)-Q-N-Q-A-W-F-S-Q-P-P-V-R-M-(3/8)和-T-Q-A-Y-K-G-F-P-V-L-P-S-(1/8),与来自阴性克隆的多肽序列-N-H-Q-S-T-F-W-Q-K-W-T-A-(6/6)比较,前3个序列在靠近肽的N端都具有相同的脯氨酸(P)和缬氨酸(V)结构(-P-V-).结论 BAC5单抗能识别近N端含有脯氨酸和缬氨酸结构的多肽.这些多肽可能模拟存在于鼻咽癌细胞表面并与BAC5单抗相关的抗原表位的构象.
目的 鑒定能被BAC5單抗識彆的定位于鼻嚥癌細胞錶麵的抗原錶位.方法 應用BAC5單抗作為靶抗體對噬菌體呈現的隨機12肽文庫進行3輪生物淘洗,用抗體捕穫和競爭試驗的夾心ELISA方法選擇和鑒定暘性噬菌體剋隆,對暘性和陰性噬菌體的外源性DNA片段進行序列分析,推導和比較由這些噬菌體所呈現的多肽氨基痠序列.結果 通過3輪生物淘洗能被抗體捕穫的噬菌體剋隆為77%(35/45).用競爭試驗從所捕穫的剋隆中測得8箇暘性剋隆.來自這8箇剋隆的噬菌體呈現三種外源多肽,即-H-Q-S-H-Y-P-Y-P-V-V-S-L-(4/8)-Q-N-Q-A-W-F-S-Q-P-P-V-R-M-(3/8)和-T-Q-A-Y-K-G-F-P-V-L-P-S-(1/8),與來自陰性剋隆的多肽序列-N-H-Q-S-T-F-W-Q-K-W-T-A-(6/6)比較,前3箇序列在靠近肽的N耑都具有相同的脯氨痠(P)和纈氨痠(V)結構(-P-V-).結論 BAC5單抗能識彆近N耑含有脯氨痠和纈氨痠結構的多肽.這些多肽可能模擬存在于鼻嚥癌細胞錶麵併與BAC5單抗相關的抗原錶位的構象.
목적 감정능피BAC5단항식별적정위우비인암세포표면적항원표위.방법 응용BAC5단항작위파항체대서균체정현적수궤12태문고진행3륜생물도세,용항체포획화경쟁시험적협심ELISA방법선택화감정양성서균체극륭,대양성화음성서균체적외원성DNA편단진행서렬분석,추도화비교유저사서균체소정현적다태안기산서렬.결과 통과3륜생물도세능피항체포획적서균체극륭위77%(35/45).용경쟁시험종소포획적극륭중측득8개양성극륭.래자저8개극륭적서균체정현삼충외원다태,즉-H-Q-S-H-Y-P-Y-P-V-V-S-L-(4/8)-Q-N-Q-A-W-F-S-Q-P-P-V-R-M-(3/8)화-T-Q-A-Y-K-G-F-P-V-L-P-S-(1/8),여래자음성극륭적다태서렬-N-H-Q-S-T-F-W-Q-K-W-T-A-(6/6)비교,전3개서렬재고근태적N단도구유상동적포안산(P)화힐안산(V)결구(-P-V-).결론 BAC5단항능식별근N단함유포안산화힐안산결구적다태.저사다태가능모의존재우비인암세포표면병여BAC5단항상관적항원표위적구상.
Objective To identify epitope relating to BAC5 mcAb, a kind of monoclonal antibody (mcAb) located on the surface of nasopha-ryngeal carcinoma (NPC) cells.Methods Using BAC5 mcAb as a selected target, the 3 rounds of biopanning to a 12 mer random peptide library (RPL) presen-ted by M13 phages were carried out. The positive M13 phage clones were chosen and confirmed with sandwich ELISA for antibodycapture and competitive assay. The exogenous DNA fragments in the positive/negative M13 phages were sequenced to deduce andcompare the order of the amino acids of exogenous peptides among the phage clones.Results 77% (35/45) of the phages eluted from the 3rd round of biopnning could be captured by BAC5 mcAb. The 3 kinds ofthe peptides were displayed by M13 phages from the 8 positive clones identified with competitive assay. The same character of "-P-V-"structure existed near N-terminus of the 3 different peptides, i.e. -H-Q-S-H-Y-P-Y-P-V-V-S-L- (4/8) -Q-N-Q-A-W-F-S-Q-P-V-R-M- (3/8) and T-Q-A-Y-K-G-F-P-V-L-P-S- (1/8) in comparison with the peptide" -N-H-Q-S-T-F-W-Q-K-W-T-A-" dis-played by M13 phages from the negative clones (6/6).Conclusion BAC5 mcAb can recognize the 3 kinds of the peptides with-P-V-structure near N-terminus. These peptides mimic thestructure of the epitope on the surface of NPC cells recognized by BAC5 mcAb.
