title: In vitro glycation of human immunoglobulin G.
abstract: BACKGROUND
  Glucose can covalently bind to the proteins by nonenzymatic process often termed glycation. Glycation of IgG is of special interest due to its possible influence on the functionality of immunoglobulins and overall immunocompetence.
  
  
  METHODS
  The glycation of IgG was studied using radioactive D-[U-14C]-glucose.
  
  
  RESULTS
  The kinetics, concentration/temperature dependence and distribution of glycation binding sites of human IgG were studied under various conditions. (a) In average, 0.7 glucose molecules were found to be bound per IgG after 90 days of incubation at 37 degrees C with normal serum IgG and glucose concentrations, and 3.1 molecules after incubation with glucose in the concentration characteristic for diabetics. (b) Incubation of the same solutions for 270 days at 4-8 degrees C resulted in binding of 0.3 and 0.8 glucose molecules per IgG, respectively. (c) After 90 days at 37 degrees C in the presence of 0.56 mol/l of glucose, IgG was glycated with averagely 47 glucose molecules per IgG and with 9 after 270 days at 4-8 degrees C. (d) At very high glucose concentration (1.67 mol/l) in concentrated IgG solution (87 g/l), the molar ratio glucose/IgG reached 41 after incubation at 60 degrees C for 10 h.
  
  
  CONCLUSIONS
  Glycation of IgG with glucose nearly resembled the first order reaction kinetics. No preferential glycation sites were found as bound glucose was equally distributed throughout F(ab)2 and Fc fragments as well as on the light and heavy immunoglobulin chains.
authors.affiliation: Institute of Immunology, Rockefellerova 2, HR-10 000 Zagreb, Croatia. avrdoljak@imz.hr
authors.forename: Anto
authors.initials: A
authors.lastname: Vrdoljak
authors.forename: Anda
authors.initials: A
authors.lastname: Trescec
authors.forename: Bojan
authors.initials: B
authors.lastname: Benko
authors.forename: Dragica
authors.initials: D
authors.lastname: Hecimovic
authors.forename: Mirjana
authors.initials: M
authors.lastname: Simic
mesh_terms: D004355:Drug Stability; D005947:Glucose / Q000737:chemistry; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G / Q000737:chemistry*; D016756:Immunoglobulins, Intravenous / Q000737:chemistry; D007202:Indicators and Reagents; D019999:Pharmaceutical Solutions; D013242:Sterilization; D013696:Temperature
chemical_list: D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous; D007202:Indicators and Reagents; D019999:Pharmaceutical Solutions; D005947:Glucose