Protein L-agarose for adsorption of autoantibodies: a potential tool for extracorporeal treatment.
This work investigated the adsorption of autoantibodies such as anti-SS-A/Ro, anti-SS-B/La, anti-Sm, and anti-dsDNA on protein L-agarose gel. In order to determine better conditions for IgG adsorption on this matrix, some buffer systems were tested. Adsorption data were analyzed using the Langmuir and Langmuir-Freundlich isotherm models. The experimental isotherms were best described by the Langmuir-Freundlich model, which indicated negative and positive cooperativities for binding in the presence of PBS and HEPES buffers, respectively. The K(d) values for phosphate buffered saline solution (PBS) and hydroxyethylpiperazine ethanesulfonic acid (HEPES) were 2.8 x 10(-7) M and 3.2 x 10(-7) M, respectively, which indicate a high affinity between IgG and the immobilized protein L. The amount of protein adsorbed per amount of protein loaded was high for anti-Sm (44%) and anti-dsDNA (46%), but low for anti-SS-B/La (9%). The amount of albumin adsorbed was lower than 0.06 mg/mL, which may remove the need for a plasma replacement solution in clinical apheresis.
Duarte
Isa Santos
IS
Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, Campinas, SP, Brazil.
Zollner
Ricardo de Lima
Rde L
Bueno
Sonia Maria Alves
SM
D000327:Adsorption; D001323:Autoantibodies / Q000032:analysis*; D001426:Bacterial Proteins / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis; D013205:Staphylococcal Protein A / Q000737:chemistry*
D001323:Autoantibodies; D001426:Bacterial Proteins; C055047:Ig L-binding protein, Peptostreptococcus; C034284:IgG Fc-binding protein, Streptococcus; D007136:Immunoglobulins; D012709:Serum Albumin; D013205:Staphylococcal Protein A