Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain |
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| Authors: | V Ghetie M A Till M A Ghetie J W Uhr E S Vitetta |
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| Affiliation: | Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235. |
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| Abstract: | A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-alpha-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue-Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disfulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected with HIV-1. |
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