Yuchi Gao1, Yan Jia1, Xin Wang1, Yuanbin Lu1, Youchao Dai1, Shiyan Yu1, Junai Zhang1, Junfa Xu2. 1. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics; Department of Clinical Immunology, Institute of Laboratory Medicine, Guangdong Medical College, Dongguan 523808, China. 2. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics; Department of Clinical Immunology, Institute of Laboratory Medicine, Guangdong Medical College, Dongguan 523808, China. *Corresponding author, E-mail: imxujunfa@163.com.
Abstract
OBJECTIVE: To prepare 5-fluorouracil (5-FU) immunogen and develop polyclonal antibodies against 5-FU. METHODS: The derivant of 5-FU (5-fluorouracil-1-yl-aceto amino acid, 5-FUAA) was synthesized, and then conjugated with bovine serum albumin (BSA) or ovalbumin (OVA) by carbodiimide (CDI) method. 5-FUAA conjugating BSA (5-FUAA-BSA) was used to immunize BALB/c mice to produce antiserum, and 5-FUAA conjugating OVA (5-FUAA-OVA) was used as coating antigen to detect the titer of the antiserum by indirect ELISA. Furthermore, specificity of the polyclonal antiserum was identified by ELISA and Western blotting. RESULTS: 5-FU derivant 5-FUAA was successfully synthesized and conjugated with BSA or OVA. Indirect ELISA showed that the titer of the antiserum from BALB/c mice immunized with 5-FUAA-BAS reached 1:1 280 000. Moreover, ELISA and Western blotting proved that the anti-serum could combine 5-FU specifically. CONCLUSION: The experiment has prepared high-specific and high-titer polyclonal antibody against 5-FU.
OBJECTIVE: To prepare 5-fluorouracil (5-FU) immunogen and develop polyclonal antibodies against 5-FU. METHODS: The derivant of 5-FU (5-fluorouracil-1-yl-aceto amino acid, 5-FUAA) was synthesized, and then conjugated with bovine serum albumin (BSA) or ovalbumin (OVA) by carbodiimide (CDI) method. 5-FUAA conjugating BSA (5-FUAA-BSA) was used to immunize BALB/c mice to produce antiserum, and 5-FUAA conjugating OVA (5-FUAA-OVA) was used as coating antigen to detect the titer of the antiserum by indirect ELISA. Furthermore, specificity of the polyclonal antiserum was identified by ELISA and Western blotting. RESULTS:5-FU derivant 5-FUAA was successfully synthesized and conjugated with BSA or OVA. Indirect ELISA showed that the titer of the antiserum from BALB/c mice immunized with 5-FUAA-BAS reached 1:1 280 000. Moreover, ELISA and Western blotting proved that the anti-serum could combine 5-FU specifically. CONCLUSION: The experiment has prepared high-specific and high-titer polyclonal antibody against 5-FU.