Rapid purification of human albumin with immunomagnetic microsphere technology.
OBJECTIVE
To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres (IMMS) based on enzyme-linked immunosorbent assay (ELISA) for the purification of human albumin from human serum.


METHODS
Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide (EDC). Finally rabbit anti-human serum albumin (HSA) antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum.


RESULTS
The result of the experiment was that the recovery of human albumin with IMMS was (86 +/- 4)%, and IMMS were reused for two other purifying cycles, the results of which were (69.0 +/- 0.6)% and (40.8 +/- 0.8)%, and the purity of the product was about 90%.


CONCLUSIONS
The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of high-purity HSA.
Wu
Ming-hui
MH
Center of New-Drug Development, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. wmh_0217@hotmail.com
Jiang
Ling-li
LL
Zeng
Fan-bo
FB
Wang
Ni-dan
ND
Tang
Lan
L
D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008863:Microspheres; D011137:Polystyrenes / Q000737:chemistry; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000276:immunology / Q000302:isolation & purification*
D011137:Polystyrenes; D012709:Serum Albumin