title: Relationship between human IgG structure and retention time in hydroxyapatite chromatography with sodium-phosphate gradient elution.
abstract: The aim of this study was to classify the retention time of recombinant human monoclonal antibodies (rhMAbs) in hydroxyapatite chromatography with sodium-phosphate gradient elution according to their physicochemical properties. We analyzed 37 rhMAbs and found that (i) the structures of both the constant and variable regions affected retention time, (ii) the number of basic amino acid residues in the variable region, particularly in the heavy chain, correlated well with retention time, and (iii) this correlation was more pronounced (e.g. r(2)=0.87 for 18 kappa IgG(1) rhMAbs) when the surface accessibility of those residues are taken into account. These findings provide a useful guide for investigators and purification-process developers working with monoclonal antibodies.
authors: Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin, Co., Ltd., Gunma, Japan. taishiro.nakagawa@kyowa-kirin.co.jp
authors: Taishiro
authors: T
authors: Nakagawa
authors: Takashi
authors: T
authors: Ishihara
authors: Hideaki
authors: H
authors: Yoshida
authors: Takashi
authors: T
authors: Yoneya
authors: Kaori
authors: K
authors: Wakamatsu
authors: Toshihiko
authors: T
authors: Kadoya
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics; D016466:CHO Cells; D002845:Chromatography / Q000295:instrumentation / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D017886:Durapatite / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000235:genetics; D010710:Phosphates / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012997:Solvents / Q000737:chemistry
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010710:Phosphates; D011134:Polysaccharides; D011994:Recombinant Proteins; D012997:Solvents; D017886:Durapatite; C018279:sodium phosphate