TẠO CỘNG HỢP VÀ ĐÁNH GIÁ CHẤT LƯỢNG KHÁNG THỂ CHUỘT KHÁNG IgG NGƯỜI GẮN ENZYME HORSERADISH PEROXIDASE ỨNG DỤNG TRONG CÁC PHƯƠNG PHÁP ELISA SỬ DỤNG KHÁNG THỂ NGƯỜI=DEVELOPENZYMET AND QUALITY ASSESSMENT MOUSE OF ANTI-HUMAN IgG ANTIBODYHORSERADISH PEROXIDASE CONJUGATES USING FOR ELISA TECHNIQUE TO DETECT HUMAN ANTIBOBY

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Tác giả: Dinh Bien Lai, Tram Nghia Thu Le, Phuong Nam Luu, Quoc Dang Quan

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học Công nghệ và Thực phẩm (Tên mới: Tạp chí Khoa học Đại học Công Thương), 2015

Mô tả vật lý: tr.17-27

Bộ sưu tập: Metadata

ID: 577041

Thêm vào giỏ

HRP (Horseradish peroxidase) has been known as an enzyme extracted from horseradish roots (Amoracia Rusticana). HRP plays an important role as a marker which most used as a common method of conjugations due to the relatively cheap, purified, fast signaling and highly specific enzyme. Furthermore, HRP involves small sizes and molecular weights having less obstructive space in associated with antigen or antibody. In immunological tests, such as ELISA, secondary antibody – horseradish peroxidase conjugates has been applied to increase specificity of enzyme in ELISA test such as anti-IgG, anti-IgM. Primary mice antibody-conjugated enzymes which also practiced for any serum sample performing any anti-antigen functions were observed. This project objects to conduct an immune response in mice with IgG antibodies which were purified from the serum of umbilical cord blood injected in different doses of 20 μg, 40 μg and 60 μg per dose of vaccine. After purified mouse antibody concentrations obtained the highest 0.495 mg per ml. The antibody anti-mouse antibody has been added in associated with the HRP enzymes. In conclusion, the antibody-antigen interaction was examined by direct and indirect ELISA and the level of community was in concentration of 0.125 mg/ml. Key words: HRP, horseradish root, immune response, IgG, IgM, ELISA, serum.HRP (Horseradish peroxidase) là một enzyme được chiết ra từ rễ cây Cải ngựa (Amoracia rusticana), HRP với vai trò chất đánh dấu được sử dụng nhiều nhất phương pháp gắn cộng hợp vì nó tương đối rẻ, tinh sạch, phát tín hiệu nhanh và có hoạt tính enzyme chuyên biệt rất cao. Ngoài ra do HRP có kích thước và trọng lượng phân tử nhỏ nên ít gây cản trở không gian khi liên kết với kháng nguyên hay kháng thể. Trong các thử nghiệm miễn dịch chẳng hạn như các phương pháp ELISA, để gia tăng độ đặc hiệu cho phản ứng, người ta thường sử dụng kháng thể (KT) thứ cấp gắn enzyme là những KT kháng loài như kháng IgM, kháng IgG,... Vai trò của cộng hợp KT chuột gắn enzyme peroxidase cũng được ứng dụng như một KT kháng loài liên kết đặc hiệu với KT sơ cấp ứng dụng cho bất kỳ mẫu huyết thanh nào kháng với một KN nào đó. Do đó, chúng tôi tiến hành gây đáp ứng miễn dịch trên chuột bằng kháng thể IgG người đã được tinh chế từ huyết thanh của máu cuống rốn với các liều tiêm khác nhau 20 µg, 40 µg và 60 µg/ liều tiêm. Sau khi tinh chế nồng độ kháng thể chuột thu được cao nhất là 0,495 mg/ml. Sau đó, cộng hợp kháng thể chuột kháng kháng thể người đã được gắn với enzyme HRP. Cuối cùng, cộng hợp được thử nghiệm bằng phương pháp ELISA trực tiếp và gián tiếp cho kết quả nồng độ cộng hợp còn giữ hoạt tính là 0,125 mg/ml. Từ khoá: HRP, cây cải ngựa, đáp ứng miễn dịch, IgG, IgM, ELISA, huyết thanh

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