Use of a bacterially expressed human factor IX light chain to develop polyclonal antibody anti-hFIX.
Hemophilia B is an X-linked recessive bleeding disorder caused by deficiency or malfunctioning of human coagulation factor IX (hFIX). Hemophilia B patients are treated at present by infusion of plasma derived hFIX which is not always efficient, because development of anti-hFIX antibodies (alloantibodies) in some cases inhibits the activity of the infused hFIX. The hFIX alloantibodies are directed against gamma-carboxyglutamic acid residues (Gla-domain) or protease domain in hFIX light chain. An epitope-containing fragment of hFIX light-chain was expressed in a T7-based Escherichia coli expression system and after purification, it was used for the immunization of rabbit to develop specific antibodies anti-hFIX. The plasma, derived from the immunized rabbit, was shown to be able to detect the normal hFIX, which indicates for the presence of a specific anti-hFIX antibody and supporting that a bacterially expressed hFIX subfragment might be able to neutralize the alloantibodies. Considering the importance of hFIX and its related investigations, both the produced hFIX antigen and its corresponding antibody will play important roles for experiments dealing with the production of hFIX and studies involved in the neutralization of the hFIX inhibitors in hFIX-related disorders and other clinical applications.
Safari
Sedigheh
S
National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
Zomorodipour
Alireza
A
Amirmozaffari
Nour
N
Daliri Choopari
Morteza
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D017123:Bacteriophage T7 / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology*; D005164:Factor IX / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology*; D015202:Protein Engineering / Q000379:methods*; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology*; D014162:Transfection / Q000379:methods
D000911:Antibodies, Monoclonal; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D005164:Factor IX