ABSTRACT
This paper describes the avidity of IgG antibody used for preparation of latex sensitized with IgG antibody (IgG-sensitized latex) and the agglutinability of IgG-sensitized latex in slide reversed passive latex agglutination (RPLA). Using immunodiffusion techniques, it was found that anti-canine C-reactive protein (CRP) sera from four rabbits immunized with canine CRP had the same antibody titers. However, the antibodies had different levels of avidity. When lattices were sensitized under the same condition with the IgG antibodies of different avidity levels separated from the above-mentioned antisera using Protein A and canine CRP-Sepharose 4B immunosorbent, these demonstrated different patterns of agglutinability in slide RPLA. The latex sensitized with IgG antibody of higher avidity demonstrated a stronger agglutinability.
Subject(s)
Antibody Affinity , Immunoglobulin G/immunology , Latex Fixation Tests/veterinary , Latex , Agglutination , Animals , C-Reactive Protein/immunology , Chromatography, Affinity , Dogs , Immune Sera/immunology , Immunodiffusion/veterinary , Immunoglobulin G/isolation & purification , Latex Fixation Tests/methods , Rabbits , Staphylococcal Protein A/immunologyABSTRACT
The canine C-reactive protein (CRP) fraction isolated from canine acute-phase serum on a phosphorylcholine-Sepharose 4B column was further subjected to Sephacryl S-300 gel filtration. A canine CRP fraction not containing IgM was then obtained. The antisera, obtained after several immunizations with this canine CRP fraction, contained nonspecific antibodies that reacted with albumin, transferrin and IgG in addition to CRP. This antiserum could be easily changed to monospecific canine CRP serum, when it was subjected to absorption for only 15 min using glutaraldehyde-insolubilized normal canine serum protein containing 3.5 micrograms ml-1 of CRP. Pure canine CRP was isolated with a recovery rate of 95% from canine acute-phase serum by affinity chromatography using specific anti-canine CRP antibody.
Subject(s)
Antibodies/immunology , C-Reactive Protein/isolation & purification , Chromatography, Affinity/veterinary , Acute-Phase Reaction/immunology , Animals , Antibodies/isolation & purification , Antibody Specificity , C-Reactive Protein/immunology , Chromatography, Affinity/methods , Chromatography, Gel/veterinary , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Immune Sera/immunology , Immunoelectrophoresis/veterinary , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Immunoglobulin M/isolation & purification , Serum Albumin/immunology , Transferrin/immunologyABSTRACT
The serum levels of C-reactive protein (CRP) produced as an inflammatory response in dogs with various disorders and surgical traumas were measured by enzyme-linked immunoabsorbent assay and slide reversed passive latex agglutination test (RPLA). The CRP levels were greatly increased 1-2 days after surgery in most of the dogs (n = 29) subjected to surgery. These levels had markedly decreased by the time the sutures were removed. In dogs with various disorders (n = 58), the serum CRP levels at first diagnosis were high in infectious diseases. In dogs from which paired serum samples were examined, the serum CRP usually showed a decrease with improvement in the condition (n = 11) or a terminal increase (n = 4) but, conversely, some showed an increase with improvement in the condition (n = 3).