New antibody purification procedure using a thermally responsive poly(N-isopropylacrylamide)-dextran derivative conjugate.
Through their specificity and affinity, antibodies are useful tools in research and medicine. In this study, we investigated a new type of chromatographic method using a thermosensitive polymer for the purification of antibodies against a dextran derivative (DD), as a model. The thermally reversible soluble-insoluble poly(N-isopropylacrylamide)-dextran derivative conjugate, named poly(NIPAAm)-DD, has been synthesized by conjugating amino-terminated poly(N-isopropylacrylamide) to a DD via ethyl-3-(3-dimethylaminopropyl)-carbodiimide. On one hand, this report describes the two steps of poly(NIPAAm)-DD conjugation and characterization. On the other hand, the poly(NIPAAm)-DD conjugate was used as a tool to purify polyclonal antibodies in serum samples from rabbits subcutaneously immunized with the derivatized dextran. Antibodies were purified and quantified by immunoenzymatic assays. Our results indicate that antibodies recognized both DD and poly(NIPAAm)-DD. In contrast, they did not bind to native poly(NIPAAm) or poly(NIPAAm) conjugated with another anionic dextran. We conclude that the conjugation of a polysaccharide to poly(NIPAAm) leads to an original and efficient chromatographic method to purify antibodies. Moreover, this novel method of purification is rapid, sensitive, inexpensive and could be used to purify various types of antibodies.
Anastase-Ravion
S
S
INVIMAT, Université Paris 13, Villetaneuse, France.
Ding
Z
Z
Pellé
A
A
Hoffman
A S
AS
Letourneur
D
D
D000178:Acrylamides / Q000737:chemistry*; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D003911:Dextrans / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D011817:Rabbits; D012680:Sensitivity and Specificity; D013696:Temperature
D000178:Acrylamides; D000906:Antibodies; D003911:Dextrans