Characterization of Monoclonal Antibody to L-Homocysteic Acid and Its Immunohistochemistry in Rat Hippocampus

• Published in: Current medical science 2005, Vol.25 (1), p.1-4
• Main Author: 刘长金 李爱 胡新武 唐明 杜以梅 张亮品 陈蕾
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Enzyme-Linked Immunosorbent Assay; Hippocampus - immunology; Hippocampus - metabolism; Homocysteine - analogs & derivatives; Homocysteine - immunology; Immunohistochemistry; L-高半胱氨酸; Rats; Rats, Sprague-Dawley; 人造单克隆抗体; 免疫化学; 海马区; 神经系统
• ISSN: 1672-0733; 2096-5230; 1993-1352; 2523-899X
• DOI: 10.1007/bf02831371

L-homocysteic acid (HCA) and other amino acids were conjugated to rat brain material(extracted rat brain protein) with glutaraldehyde to form HCA- and amino acids-brain material conjugates. The specificity of monoclonal antibody (McAb) was tested on serial dilution test and absorption test on enzyme-linked immunosorbent assay (ELISA) using these conjugates as antigens instead of amino acids-BSA (bovine serum albumin) conjugates used previously. The characterized McAb was applied for immunohistochemical staining using PAP (peroxidase antiperoxidase) technique in combination with silver enhancement of diamino-benzene (DAB) products. The results indicated that McAb to L-HCA reacted with L-HCA-brain material conjugates, but not with other amino acids-brain material conjugates so far tested. McAb absorbed with L-HCA-brain material abolished or decreased immunoreactivity of L-HCA-brain material with McAb. The antibody selectively stained subpopulation of cells and processes in the hippocampus fixed with glutaradehyde.Absorption of McAb with L-HCA-brain material abolished immunohistochemical staining. These results suggested that McAb was specific for L-HCA-brain materials and could be used for imunohistocytochemistry. This would provide a new tool for immunohistochemical visualization and localization of L-HCA in the nervous system.