Stability and cytotoxicity of Fab-ricin A immunotoxins prepared with water soluble long chain heterobifunctional crosslinking agents.
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]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.
Woo
B H
BH
College of Pharmacy, SungKyunKwan University, Jangan-ku, Suwon City, Korea.
Lee
J T
JT
Park
M O
MO
Lee
K R
KR
Han
J W
JW
Park
E S
ES
Yoo
S D
SD
Lee
K C
KC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002470:Cell Survival / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry*; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay; D005978:Glutathione / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry*; D000922:Immunotoxins / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D015260:Neprilysin / Q000276:immunology*; D012276:Ricin / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D012995:Solubility; D013388:Succinimides / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D004220:Disulfides; D007140:Immunoglobulin Fab Fragments; D000922:Immunotoxins; D013388:Succinimides; D012276:Ricin; D015260:Neprilysin; D005978:Glutathione