高等学校化学学报

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (4): 530.

• 研究论文 • 上一篇    下一篇

抗meso-四(α,α,α,α-O-苯乙酰苯)卟啉抗体的酶学性质研究

祁超1,2, 丁兰1, 丁天兵3, 刘志强1, 张玉静2, 刘立岩1, 赵大庆1   

  1. 1. 中国科学院长春应用化学研究所稀土化学与物理开放实验室, 长春 130022;
    2. 中国人民解放军军需大学基础部生物化学教研室, 长春 130062;
    3. 吉林大学分子酶学工程教育部开放实验室, 长春 130023
  • 收稿日期:2001-05-09 出版日期:2002-04-24 发布日期:2002-04-24
  • 通讯作者: 赵大庆(1963年出生),男,博士,研究员,主要从事生物无机化学研究.
  • 基金资助:

    国家自然科学基金(批准号:29701005);中国科学院基础局重大项目(批准号:KJ951－A1－504－02)资助.

Studies on Enzymological Properties of Antibody Elicited bymeso-Tetra(α, α, α, α-O-phenylacetyl benzene)porphyrin

QI Chao1,2, DING Lan1, DING Tian-Bing3, LIU Zhi-Qiang1, ZHANG Yu-Jing2, LIU Li-Yan1, ZHAO Da-Qing1   

  1. 1. Key Laboratory of Rare Earth Chemistry and Physics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
    2. Biochemistry Section of Basic Department, Quartermaster University of PLA, Changchun 130062, China;
    3. Key Laboratory of Molecular Enzymology and Engineering of Educational Ministry, Jilin University, Changchun 130023, China
  • Received:2001-05-09 Online:2002-04-24 Published:2002-04-24

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摘要： 利用小分子meso-四(α,α,α,α-O-苯乙酰苯)卟啉作为全抗原免疫小鼠,通过细胞融合等技术制备了单克隆抗体(McAb)1F2,利用高效液相色谱(HPLC)、基质辅助激光解吸飞行时间质谱(MALDI/TOFMS)证明纯化得到的单抗是很纯的。McAb1F2的保留时间是2.63min,亚型为IgG2a,相对分子量为156678.8.McAb1F2-卟啉复合物形成时,卟啉Soret带最大吸收峰从408nm红移到416nm,并有增色效应,说明McAb1F2-卟啉是刚性且紧密结合。抗体酶稳定性很高,但比活力只有4.6875U/mg,为天然酶HRP的1.899%.抗体酶的Km=20.29mmol/L,kcat=396.82min-1,kcat/Km=1.9557×104L·mol-1·min-1.

关键词: meso-四(&alpha, &alpha, &alpha, &alpha, -O-苯乙酰苯)卟啉, 单克隆抗体, 全抗原, 过氧化物酶

Abstract: meso-Tetra(α,α,α,α-O-phenylacetyl benzene)porphyrin was used as a complete antigen to elicit monoclonal antibody 1F2 through the immunization and cell fusion techniques.McAb1F2 obtained was demonstrated very pure by HPLCand MALDI/TOFMS.The retention time of McAb1F2 was 2.63 min.The subtype of McAb1F2 was IgG2a.The relative molecular weight was 156678.8.When the McAb1F2-porphyrin was formed, the maximal absorption of the porphyrin soret region had a redshift from 408 to 416 nm and hyperchromical effect, showing that the antigen-antibody combination was rigid and intense , and the abzyme constancy was high.But compared with HRP, the activity of the abzyme was only 4.6875 U/mg and 1.899% of that of HRP.Its Km was 20.29 mmol/L, kcat 396.82 min-1, kcat/Km 1.9557×104 L·mol-1·min-1.

Key words: meso-Tetra (α,α,α,α-O-phenylacetyl benzene)porphyrin, McAb, Complete antigen, Peroxidase

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