Obtaining of the heavy chains of the human IgG for the preparation of a specific immunoserum for their use in the immunodiagnosis

Antonio Melchor-Rodríguez; Vivian Alonso-Ramos; Raquel López-Cisnero; Orlando Zulueta-Rodríguez; Carlos Martínez-Ortiz; Liliana Hernández-Pérez; Ivonne Gómez-Cordero; Liliena López-Brauet

2002, Number 4

Bioquimia 2002; 27 (4)

Obtaining of the heavy chains of the human IgG for the preparation of a specific immunoserum for their use in the immunodiagnosis

Antonio Melchor-Rodríguez, Vivian Alonso-Ramos, Raquel López-Cisnero,Orlando Zulueta-Rodríguez1, Carlos Martínez-Ortiz, Hernández-Pérez L, Gómez-Cordero I, López-Brauet L

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ABSTRACT

The method is described for purification of the heavy chain of immunoglobulin G (IgG) from a pool of normal human serum. The IgG was purified by ammonium sulphate precipitation and ion-exchange chromatography on DEAE-Sephacel. The purified preparation was homogeneous by sodium dodecylsulphate polyacrylamide (SDS-PAGE) and tested by immunoelectrophoresis. The final recovery of human IgG was 68.78 % and a purification factor of 4.01. The heavy chains of IgG were obtained by reduction with dithiothreitol and alkylation with iodoacetamide. The separation of heavy and light chains of IgG was done by gel filtration on Sephacryl S-100 HR in two columns connected in series. The obtained preparation was homogeneous by double immunodiffusion and SDS-PAGE. The relative molecular weight of heavy chains of IgG was 58.5 kDa. The yield was 0.603 mg of heavy chains per mg of IgG.

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