Allergen-Specific IgG Antibodies Purified from Mite-Allergic Patients Sera Block the IgE Recognition of Dermatophagoides pteronyssinus Antigens: An In Vitro Study
Figure 3
(a) Representative chromatogram of the total human IgG purification by affinity chromatography in protein G-agarose obtained from the 40% ammonium sulfate precipitated fraction (P 40%) of a serum pool. I—elution peak of P 40% nonligand fraction after washing with 0.02 M phosphate buffer pH 8.0 (black arrow); II—elution peak of P 40% ligand fraction after washing with 0.1 M glycine buffer pH 2.6 (dashed arrow). Data are expressed in absorbance (280 nm). Elution volume consisted of 1 mL in each tube. Values of pH were also measured in each elution tube. (b) Electrophoretic profile in SDS-PAGE 8% stained with blue silver. NLF—nonligand fraction, LF—ligand fraction, corresponding to the tubes 29–34. Markers of molecular weight (MW) are indicated on the left in kilodaltons (kDa). (c) Immunoblots for detection of IgG, IgA, IgE, and IgM in the serum fractions after purification in protein G-Agarose as shown in (b). Bands were revealed with DAB as described in Methods.
