A method for studying insoluble immune complexes.
Two variants of a method for determining the average composition of insoluble immune complex particles (IICP) are described. The first variant is based on measuring the specific turbidity (the turbidity per unit mass concentration of the dispersed substance) and the average size of IICP determined from dynamic light scattering (DLS). In the second variant, the slope of the logarithmic turbidity spectrum (wavelength exponent) is used instead of DLS particle size. Both variants allow the average biopolymer volume fraction to be determined in terms of the average refractive index of IICP. The method is exemplified by two experimental antigen+antibody systems: (i) lipopolysaccharide-protein complex (LPPC) of Azospirillum brasilense Sp245+rabbit anti-LPPC; and (ii) human IgG (hIgG)+sheep anti-hIgG. We have found that IICP can be modeled by incompact porous particles that contain about 30% of biopolymer substance and 70% of buffer.
Khlebtsov
Boris N
BN
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, 13 Pr. Entuziastov, Saratov 410049, Russia. khlebtsov@ibppm.sgu.ru
Burygin
Gennadii L
GL
Matora
Larisa Yu
LY
Shchyogolev
Sergei Yu
SY
Khlebtsov
Nikolai G
NG
D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000737:chemistry*; D015806:Azospirillum brasilense; D011232:Chemical Precipitation; D003102:Colloids / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D008027:Light; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology; D008962:Models, Theoretical; D009391:Nephelometry and Turbidimetry / Q000379:methods*; D010316:Particle Size; D012031:Refractometry / Q000379:methods*; D012542:Scattering, Radiation; D012995:Solubility; D013053:Spectrophotometry
D000888:Antibodies, Anti-Idiotypic; D000936:Antigen-Antibody Complex; D003102:Colloids; D007074:Immunoglobulin G; D008070:Lipopolysaccharides; C048844:anti-IgG