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Design and optimization of primer for the cloning of the mouse Fd immunoglobulin M for antibody phage display technology |
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| รหัสดีโอไอ | |
| Creator | Sitthinon Siripanthong |
| Title | Design and optimization of primer for the cloning of the mouse Fd immunoglobulin M for antibody phage display technology |
| Contributor | Achara Phumyen, Anchalee Techasen, Paiboon Sithithaworn, Prasert Saichua, Chanvit Leelayuwat, Amonrat Jumnainsong |
| Publisher | Faculty of Associated Medical Sciences, Khon Kaen University, Thailand |
| Publication Year | 2564 |
| Journal Title | Archives of Allied Health Sciences |
| Journal Vol. | 33 |
| Journal No. | 1 |
| Page no. | 70-78 |
| Keyword | Opisthorchis viverrini, Antibody, Phage display technology, Primer |
| URL Website | https://he01.tci-thaijo.org/index.php/ams/about |
| Website title | Archives of Allied Health Sciences (Arch AHS) |
| ISSN | 2730-1990 |
| Abstract | Hybridoma technique is worldwide used for antibody production. However, there are many issues of using this technique such as hybridoma instability, chromosome loss, mutation and short haft-life that lead to loss of hybridoma genome and loss of antibody production. The technique which displays antibody on phage particle can solve this problem. Therefore, antibody phage display was used to sustain the hybridoma technique to produce antibodies. In this technique, it is needed to amplify antibody sequence by a primer that is specifc to antibody isotype and have cloning site to clone antibody sequence into a phagemid vector. Unfortunately, there was no reverse primer for amplifying mouse Fd immunoglobulin M (IgM) and clone into the pComb3HSS phagemid vector. The aim of this study was to design the new reverse primers to amplify mouse Fd immunoglobulin M antibody gene by polymerase chain reaction (PCR) using KKU505 hybridoma as a model. The KKU505 hybridoma can produce anti-Opisthorchis viverrini (O. viverrini) monoclonal antibody (mAb). The results demonstrated that this new primer could amplify the Fd immunoglobulin M of anti-Opisthorchis viverrini mAb and construct into pComb3HSS phagemid vector. This technology could preserve antibody gene and use for more stable antibody production. |
