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KMID : 0043319990220050459
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Archives of Pharmacal Research
1999 Volume.22 No. 5 p.459 ~ p.463
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Stability and Cytotoxicity of Fab-Ricin A Immunotoxins Prepared with Water Soluble Long Chain Heterobifunctional Crosslinking Agents
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Woo Byung-Ho
Lee Jung-Tae
Park Myung-Ok
Lee Kang-Ro
Han Jeung-Whan
Park Eun-Seok
Yoo Sun-Dong
Lee Kang-Choon
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Abstract
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The effects of the hindered and non-hindered water soluble long-chain disulfide bonds on the stability and cytotoxicity of the ricin A chain (RTA) immunotoxin were examined. The RTA immunotoxins were prepared with the Fab fragments of anti-common acute lymphoblastic leukemia antigen (CALLA) monoclonal antibody (Fab-RTA) using sulfosuccinimidyl-6-[(-methyl-(-2-pyridyldithio)toluamido]toluamido]hexanoate (S-LC-SMPT) and sulfosuccinimidyl-6-[3-(2-pyridyldithio-propionamido]hexanoate (S-LC-SPDP). The prepared Fab-RTA immunotoxins were evaluated for their conjugation yield, immunoreactivity, thermal and disulfide bond stability and cytotoxicity. The conjugation yield of the Fab-RTA immunotoxin from the water soluble long chain crosslinking agents, S-LC-SMPT and S-LC-SPDP, were comparable. Both Fab-RTA immunotoxins exhibited a similar immunoreactivity and thermal stability in aqueous solution. However, S-LC-SMPT -mediated Fab-RTA, sterically hindered, showed an enhanced disulfide bond stability in vitro over S-LC-SPDP mediated one. In the cytotoxicity against antigenic cell Daudi, the S-LC-SMPT -mediated RTA immunotoxin maintained a comparable cytotoxicity, compared with S-LC-SPDP mediated Fab-RTA immunotoxin.
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KEYWORD
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Immunotoxin, Ricin A chain, Fab, Monoclonal antibody, S-LC-SMPT, S-LC-SPDP
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