Production and purification of ovine anti-tetanus antibody.
We used the ovine as bioreactor for the production and optimization of anti-tetanus toxin antibody. Four female sheep were immunized with human tetanus vaccine (TT-alum) every two weeks for 16 weeks, after which serum was collected and its titer was estimated by ELISA. The highest titer obtained was 39,000 IU ml-1. To optimize a purification protocol for ovine anti-tetanus toxin, we used four procedures; weak anion (DEAE-Sephadex), weak cation (CM-Sephadex), ammonium sulfate precipitation alone or in combination with caprylic acid. Fifty percent saturation with ammonium sulfate combined with caprylic acid gave us the highest yield of protein with specific activity and the purest Fab product.
Redwan
El-Rashdy M
el-RM
Department of Protein Research, GEBRI, Mubarak City for Scientific Research and Biotechnology Applications, New Borg EL-Arab, Alexandria 71787, Egypt. redwan1961@yahoo.com
Khalil
Ahmed
A
El-Dardiri
Zeinab Z
ZZ
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000918:Antibody Specificity; D019149:Bioreactors; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange / Q000662:veterinary; D005260:Female; D006801:Humans; D007114:Immunization / Q000662:veterinary*; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000276:immunology; D012756:Sheep / Q000276:immunology*; D013742:Tetanus / Q000517:prevention & control*; D013743:Tetanus Antitoxin / Q000096:biosynthesis* / Q000097:blood / Q000737:chemistry / Q000494:pharmacology; D013745:Tetanus Toxoid / Q000276:immunology / Q000302:isolation & purification*
D000907:Antibodies, Bacterial; D007140:Immunoglobulin Fab Fragments; D013743:Tetanus Antitoxin; D013745:Tetanus Toxoid