Optimization of Conditions for Formation and Analysis of Anti-CD19 FVS191 Single-Chain Fv Homodimer (scFv')2
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- 作者：Duo Wang ; Erica Berven ; Quanzhi Li ; Fatih Uckun ; and John H. Kersey
- 刊名：Bioconjugate Chemistry
- 出版年：1997
- 出版时间：January 1997
- 年：1997
- 卷：8
- 期：1
- 页码：64 - 70
- 全文大小：161K
- 年卷期：v.8,no.1(January 1997)
- ISSN：1520-4812
文摘
In this report, we present the production of a dimeric form of anti-CD19scFv, the FVS191cys (scFv')2.Anti-CD19 scFv FVS191cys was constructed by engineering a cysteineresidue at the C terminus ofthe VL domain of scFv FVS191. FVS191cys(scFv')2 was formed through a disulfide bondbetweentwo FVS191cys molecules. To optimize the yield of FVS191cys(scFv')2, the effects of oxidation time,buffer pH, and temperature on the formation of dimeric scFv wereanalyzed. Our study indicatesthat the formation of FVS191cys (scFv')2 is oxidation time-and buffer pH-dependent; a high pH bufferfacilitates the formation of disulfide-linked (scFv')2.The maximum yield of FVS191cys (scFv')2canbe achieved when FVS191cys is air-oxidized at 4 
C, in buffer with apH of 8.5-9. The biologicalactivity of FVS191cys (scFv')2 was analyzed by ELISA and aninternalization assay. FVS191cys (scFv')2has a CD19 binding ability similar to that of its parental mAb B43 andis internalized by CD19 positiveNalm 6 cells. This study indicates that FVS191cys(scFv')2 is a potential candidate for tumordiagnosisor therapy.
C, in buffer with apH of 8.5-9. The biologicalactivity of FVS191cys (scFv')2 was analyzed by ELISA and aninternalization assay. FVS191cys (scFv')2has a CD19 binding ability similar to that of its parental mAb B43 andis internalized by CD19 positiveNalm 6 cells. This study indicates that FVS191cys(scFv')2 is a potential candidate for tumordiagnosisor therapy.