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2.63 min,亚型为IgG2a,相对分子量为156 678.8.McAb 1F2-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4.687 5 U/mg,为天然酶HRP的1.899%.抗体酶的Km=20.29 mmol/L,kcat=396.82 min-1,kcat/Km=1.955 7×104L·mol-1·min-1.
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. McAb 1F2 obtained was demonstrated very pure by HPLC and MALDI/TOFMS. The retention time of McAb 1F2 was 2. 63 min. The subtype of McAb 1F2 was IgG2a. The relative molecular weight was 156 678. 8. When the McAb 1F2-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. 687 5 U/mg and 1. 899% of that of HRP. Its Km was 20. 29 mmol/L, kcat 396. 82 min-1, kcat/Km 1. 955 7×104 L.mol-1.min-1.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2002年第4期530-534,共5页
Chemical Journal of Chinese Universities
基金
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.