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KMID : 0390320170270020007
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Chungbuk Medical Journal
2017 Volume.27 No. 2 p.7 ~ p.15
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Establishment of CEACAM6 Sandwich ELISA for Diagnosis of Solid Tumor
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Kim Min-Young
Seon Seong-Gyu
Park Bo-Reum
Song Hyung-Geun
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Abstract
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Purpose: CEACAM6 is a 90-kDa GPI-linked glycoprotein, found at relatively high levels in serum from lung, pancreatic, breast, and colorectal cancer patients. The purpose of this study was to establish a CEACAM6 specific sandwich ELISA for measuring the amount of CEACAM6 in the blood of cancer patients.
Materials and Methods: CEACAM6 gene was isolated from a non-small cell lung cancer cell line A549 by PCR and recombined with an expression vector (pSecTag) designed to express together with the constant region of human antibody through gene cloning. The recombinant expression vector was expressed in HEK-293T cells and the reactivity of the produced recombinant CEACAM6 was confirmed by flow cytometry and ELISA using 8F5, 9A6 and 18B8 known as anti-CEACAM6 antibodies.
Results: Approximately 1 kb of CEACAM6 gene was obtained from non-small cell lung cancer cell line A549, and recombinant CEACAM6 fused with human antibody constant region were developed. The standard curve of the CEACAM6 specific sandwich ELISA was established using recombinant CEACAM6 and anti-CEACAM6 antibodies 8F5, 9A6 and 18B8.
Conclusion: Through this study, CEACAM6 specific sandwich ELISA was established to quantify CEACAM6 in patient peripheral blood.
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KEYWORD
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Non-small cell lung cancer, CEACAM6, ELISA
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