Rapid fractionation of serum immunoglobulins by high pressure liquid gel permeation chromatography. Application to routine serologic procedures.
High pressure liquid chromatography (HPLC) was applied to separation of IgM from IgG and IgA for the detection of virus-specific antibodies by routine serologic methods. Serum samples of 250 microliter were fractionated on a HPLC protein column after filtration for use in rubella haemagglutination inhibition after preabsorption with kaolin. The protein fractions were examined for IgM, IgG and IgA content and for cross contamination. The relative recovery after kaolin absorption was satisfactory: IgM greater than 60%, IgG greater than 90%, IgA greater than 90%. The immunoglobulin M was well separated. IgG/IgA contamination of the fraction was less than 0.2%/ less than 0.5%. The IgM and IgG fractions were used without further treatment in haemagglutination inhibition, complement fixation and enzyme-linked immunosorbent assay. Highly specific results in the diagnosis of acute viral diseases (Rubella, Herpes zoster, cytomegalovirus) and Mycoplasma pneumoniae were obtained.
D000907:Antibodies, Bacterial / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D001769:Blood / Q000382:microbiology; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003168:Complement Fixation Tests; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D014777:Virus Diseases / Q000097:blood
D000907:Antibodies, Bacterial; D000914:Antibodies, Viral; D007075:Immunoglobulin M