International Journal of Scientific and Research Publications

IJSRP, Volume 3, Issue 4, April 2013 Edition [ISSN 2250-3153]

Preparation of Stable and Highly Active Hemoglobin by Using Antiglycosylspecific Hemoglobin Polyclonal Antibodies
      Jan. U, Parveen.S, Hussain.Q
Abstract: The highly purified oligosaccharide chains obtained from glycoprotein invertase were coupled with bovine hemoglobin with the help of cross linking agent .The neoglycoprotein hemoglobin was highly purified to homogeneity by column chromatography and characterized on SDS- PAGE . Rabbits were immunized to produce glycosylspecific antihemoglobin polyclonal antibodies. Antibodies were purified by (NH4)SO2 precipitation followed by DEAE cellulose chromatography. The IgG-Sepharose was prepared by covalently coupling the purified polyclonal antibodies to CNBr-activated Sepharose 4B. The large amount of hemoglobin was immobilized on IgG-Sepharose by alternate incubation of hemoglobin and glycosylspecific antihemoglobin polyclonal IgG. The immunoaffinity based layered assembled preparation were highly active .The amount of hemoglobin immobilized could be raised 55 fold after four binding cycles. A layer by layer immobilization of hemoglobin significantly increases its life span and improved in stability against high temperature and other several denaturants (pH and salicylate). The results show the excellent platform for protein immobilization with high affinity. The polyclonal antibodies as supporting material has been found challenging for immobilizing the proteins. Further antiglycosylated hemoglobin can be used as a nanoparticle for immobilizing high amount of hemoglobin with no cytotoxicity to cell and this technique enhances the incorporation of hemoglobin properties (recognition, transport and catalytic properties). The applications of the immobilized glycosylated hemoglobin are found in the field of medicine, biotechnology, nanotechnology and biosensors.

Reference this Research Paper (copy & paste below code):

Jan. U, Parveen.S, Hussain.Q (2018); Preparation of Stable and Highly Active Hemoglobin by Using Antiglycosylspecific Hemoglobin Polyclonal Antibodies; Int J Sci Res Publ 3(4) (ISSN: 2250-3153).
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