[Studies of the expression, purification, renaturation and biologic activity of an anti-CEA immunotoxin].
A recombinant immunotoxin named CEA/PE38/KDEL was constructed, which was composed of anti-CEA single-chain Fv and the truncated and modified form of Pseudomonas exotoxin (PE38/KDEL). The CEA/PE38/KDEL immunotoxin was expressed in the E. coli strain BL21 (DE3)-star as inclusion bodies. The denatured inclusion bodies were purified with Ni-NTA chelate agarose, then the constant gradient dialysis was used to perform the refolding of the CEA/PE38/KDEL immunotoxin. Results of FACS and MTT assay indicate that the refolded immunotoxins keep potent and specific cytotoxicity to tumor cells bearing CEA antigens.
Yang
Hui
H
School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110015, China.
He
Dan
D
Chao
Kai
K
Lin
Qing
Q
You
Song
S
Huang
Hua-Liang
HL
D036002:ADP Ribose Transferases / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D000906:Antibodies / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000378:metabolism* / Q000494:pharmacology; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005098:Exotoxins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics; D000922:Immunotoxins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D020673:Protein Renaturation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D037521:Virulence Factors / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D000906:Antibodies; D000970:Antineoplastic Agents; D001427:Bacterial Toxins; D002272:Carcinoembryonic Antigen; D005098:Exotoxins; D007128:Immunoglobulin Fragments; D000922:Immunotoxins; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A; C055966:immunoglobulin Fv