title: Affinity purification of antibodies by using Ni2+-resins on which inclusion body-forming proteins are immobilized.
abstract: Bacterially expressed recombinant proteins are widely used for producing specific antibodies. Unfortunately, many recombinant proteins are recovered as insoluble materials, so-called inclusion bodies. Inclusion bodies are rather advantageous from a point of view of immunogens because fairly pure proteins can be feasibly extracted from the inclusion bodies. However, we encounter a problem with an insoluble protein when we make an antigen-immobilized column for affinity purification of antibodies because we need a soluble protein in usual immobilization methods. Histidine-tagged proteins can be bound to Ni(2+)-resins in buffer containing 6M guanidine-HCl, in which most insoluble proteins are solubilized. Taking advantage of this feature, we have successfully purified antigen-specific antibodies by directly using Ni(2+)-resins onto which denatured proteins are bound.
authors.affiliation: Department of Clinical Chemistry and Laboratory Medicine, Kyushu University Graduate School of Medical Sciences, Fukuoka 812-8582, Japan.
authors.forename: Worawan
authors.initials: W
authors.lastname: Chumpia
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohsato
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kuma
authors.forename: Shogo
authors.initials: S
authors.lastname: Ikeda
authors.forename: Naotaka
authors.initials: N
authors.lastname: Hamasaki
authors.forename: Dongchon
authors.initials: D
authors.lastname: Kang
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D002460:Cell Line; D002846:Chromatography, Affinity; D045303:Cytochromes b / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry*; D009532:Nickel / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012995:Solubility
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D009532:Nickel; D045303:Cytochromes b; D004706:Endodeoxyribonucleases; C052391:endonuclease G