DESIGN OF SUPER CATALYTIC ANTIBODIES DESTROYING TARGETING VIRUS AND BACTERIUM

通过免疫基态肽或蛋白质，我们可以产生具有丝氨酸蛋白酶样特征的“超催化抗体”。在本研究中，我们成功制备了i41-7、i41SL-1-2、幽门螺杆菌脲酶和趋化因子受体CCRS肽等“超催化抗体”。(i)i41-7我们制备了6种针对亲本41S-2 mAb的抗独特型单克隆抗体（mAb），其轻链是一种能够降解靶向HIV-1gp41分子的超催化抗体（41S-2- l）。轻链和重链具有由Ser、His和Asp三种构象组成的催化三联结构。正如预期的那样，i41-7单抗的两条链都可以切割一些肽的肽键，如多肽TP41-1 （TPRGPDRPEGIEEEGGERDRD）。（ii）通过免疫超催化抗体轻链41S-2-L的互补决定区-1肽（CDRL-1: RSSKSLLYSNGNTYLY）获得i4LSL1-2单克隆抗体（mAb） i4LSL1-2，该单克隆抗体能够更有效地破坏HIV-1包膜的gp41分子。i41SL1-2 - i41SL1-2单抗的轻链和重链在其结构上具有催化三联体。i41SL1-2单抗的轻链和重链分别在47和57小时内降解抗原肽CDRL-1。利用合成的HpU-2表位序列SVELIDIGGNRRIFGFNALVDR对HpU-2轻链、9和18的催化活性进行了研究。轻链降解表位肽呈双相反应。此外，它对幽门螺杆菌脲酶有破坏作用，对牛血清白蛋白无破坏作用。（IV）趋化因子受体CCR5A单克隆抗体ECL2B-2通过免疫含有趋化因子受体CCR-5部分序列的肽获得，该受体作为膜蛋白存在于巨噬细胞表面，在HIV感染中起重要作用。ECL2B-2单抗轻链在约100小时内降解抗原肽CCR-5。令人惊讶的是，轻链具有非常高的催化反应速率常数（kcat），为2.23 min^<-1>，比以前获得的天然催化抗体大几十到几百倍。少

By immunizing ground-state peptides or proteins, we can produce "super catalytic antibodies" possessing serine protease-like characteristics. In this study, we succeeded in preparing "super catalytic antibodies" such as i41-7, i41SL-1-2, Helicobacter pylon urease and chemokine receptor CCRS peptide, and (iii) others.(i)i41-7We prepared six anti-idiotypic monoclonal antibodies (mAbs) against parent 41S-2 mAb whose light chain is a super catalytic antibody (41S-2-L) capable of degrading targeted HIV-1gp41 molecule. The light and heavy chain possess catalytic triad-like structure composed of Ser, His and Asp in their conformations. Both chains of i41-7 mAb could cleave peptide bond of some peptides such as a polypeptide, TP41-1 (TPRGPDRPEGIEEEGGERDRD), as anticipated.(ii)i41SL-1-2A monoclonal antibody (mAb) i4LSL1-2 was obtained by immunizing the peptide of complementarity determining region-1 (CDRL-1: RSSKSLLYSNGNTYLY) of super catalytic antibody light chain, 41S-2-L, capable of emzymati … More cally destroying the gp41 molecule of HIV-1 envelope. The light and heavy chain of i41SL1-2 i41SL1-2 mAb possess catalytic triads in their structures. Both light and heavy chains of i41SL1-2 mAb degraded the antigenic peptide CDRL-1 within 47 and 57 hr, respectively.(iii)Helicobacter pyroni ureaseThe catalytic activities of the light chain of HpU-2,9,and 18 were investigated using the synthetic peptide SVELIDIGGNRRIFGFNALVDR which is the epitope sequence of HpU-2. The light chains could degrade the epitope peptide as showing biphasic reaction profile. Moreover, it destroyed H.pyloni urease but not BSA.(IV)Chemokine receptor CCR5A monoclonal antibody (mAb), ECL2B-2, was obtained by immunizing a peptide possessing a part of a sequence of a chemokine receptor, CCR-5, which is present as a membrane protein on the surface of macrophage and which plays an important role in HIV infection. The light chain of ECL2B-2 mAb degraded the antigenic peptide CCR-5 within about 100 hr. Surprisingly, the light chain had a very high catalytic reaction rate constant (kcat) of 2.23 min^<-1>, which is greater by factors of tens to hundreds than those of natural catalytic antibodies previously obtained. Less