Preparation of rabbit monoclonal antibody against cGMP and development of competitive ELISA for cGMP / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 423-428, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-255174
ABSTRACT
<p><b>OBJECTIVE</b>To prepare rabbit monoclonal antibody (RabMab) against guanosine 3', 5'-cyclic monophosphate (cGMP) and to develop a competitive ELISA for the detection of cGMP.</p><p><b>METHODS</b>New Zealand white rabbits were immunized with synthesized cGMP-keyhole limpet hemoeyanin (cGMP-KLH) to prepared a RabMAb with monoclonal antibody technique of Epitomics. A competitive ELISA kit was produced with cGMP RabMAb. The specificity, the precision and the recoveries of the method were determined.</p><p><b>RESULTS</b>The RabMAb with high sensitivity towards cGMP were prepared with an antibody timer of 3.1 ng/mL and 50% inhibitive concentration (IC50) of 12.57 ng/mL. The cGMP RabMAb had 33% cross-reactivity to inosine 3', 5'-cyclic monophosphate (cIMP) and little or no cross-reactivity to other compounds. A competitive ELISA was developed for detection of cGMP. The range of detection was 0~120 ng/mL with a minimal limit of 1.95 ng/mL. The recovery of assay was 89%~103%. The inter-assay and intra-assay coefficient variations were below 11.68% and 13.85%, respectively.</p><p><b>CONCLUSION</b>The RabMab against cGMP with high affinity and high specificity has been generated successfully, and a competitive ELISA for detection of cGMP has been developed with the prepared cGMP RabMAb.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Enzyme-Linked Immunosorbent Assay
/
Cyclic GMP
/
Cross Reactions
/
Allergy and Immunology
/
Antibodies, Monoclonal
/
Antibody Specificity
Limits:
Animals
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2015
Type:
Article
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