Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity
<em>Naja kaouthia</em> (monocled cobra) venom contains many isoforms of secreted phospholipase A2 (sPLA<sub>2</sub>). The PLA<sub>2</sub> exerts several pharmacologic and toxic effects in the snake bitten subject, de...
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| Format: | Article |
| Language: | English |
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MDPI AG
2012-07-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/4/7/554 |
| _version_ | 1817994339250339840 |
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| author | Wanpen Chaicumpa Nitat Sookrung Kunan Bangphoomi Charnwit Chavanayarn Jeeraphong Thanongsaksrikul Kanyarat Thueng-in |
| author_facet | Wanpen Chaicumpa Nitat Sookrung Kunan Bangphoomi Charnwit Chavanayarn Jeeraphong Thanongsaksrikul Kanyarat Thueng-in |
| author_sort | Wanpen Chaicumpa |
| collection | DOAJ |
| description | <em>Naja kaouthia</em> (monocled cobra) venom contains many isoforms of secreted phospholipase A2 (sPLA<sub>2</sub>). The PLA<sub>2</sub> exerts several pharmacologic and toxic effects in the snake bitten subject, dependent or independent on the enzymatic activity. <em>N. kaouthia</em> venom appeared in two protein profiles, P3 and P5, after fractionating the venom by ion exchange column chromatography. In this study, phage clones displaying humanized-camel single domain antibodies (VH/V<sub>H</sub>H) that bound specifically to the P3 and P5 were selected from a humanized-camel VH/V<sub>H</sub>H phage display library. Two phagemid transfected <em>E. coli</em> clones (P3-1 and P3-3) produced humanized-V<sub>H</sub>H, while another clone (P3-7) produced humanized-VH. At the optimal venom:antibody ratio, the VH/V<sub>H</sub>H purified from the <em>E. coli</em> homogenates neutralized PLA<sub>2</sub> enzyme activity comparable to the horse immune serum against the <em>N. kaouthia</em> holo-venom. Homology modeling and molecular docking revealed that the VH/V<sub>H</sub>H covered the areas around the PLA<sub>2</sub> catalytic groove and inserted their Complementarity Determining Regions (CDRs) into the enzymatic cleft. It is envisaged that the VH/V<sub>H</sub>H would ameliorate/abrogate the principal toxicity of the venom PLA<sub>2</sub> (membrane phospholipid catabolism leading to cellular and subcellular membrane damage which consequently causes hemolysis, hemorrhage, and dermo-/myo-necrosis), if they were used for passive immunotherapy of the cobra bitten victim. The speculation needs further investigations. |
| first_indexed | 2024-04-14T01:51:34Z |
| format | Article |
| id | doaj.art-6d317906c16a4fdab170326561459a7a |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-14T01:51:34Z |
| publishDate | 2012-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-6d317906c16a4fdab170326561459a7a2022-12-22T02:19:20ZengMDPI AGToxins2072-66512012-07-014755456710.3390/toxins4070554Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic ActivityWanpen ChaicumpaNitat SookrungKunan BangphoomiCharnwit ChavanayarnJeeraphong ThanongsaksrikulKanyarat Thueng-in<em>Naja kaouthia</em> (monocled cobra) venom contains many isoforms of secreted phospholipase A2 (sPLA<sub>2</sub>). The PLA<sub>2</sub> exerts several pharmacologic and toxic effects in the snake bitten subject, dependent or independent on the enzymatic activity. <em>N. kaouthia</em> venom appeared in two protein profiles, P3 and P5, after fractionating the venom by ion exchange column chromatography. In this study, phage clones displaying humanized-camel single domain antibodies (VH/V<sub>H</sub>H) that bound specifically to the P3 and P5 were selected from a humanized-camel VH/V<sub>H</sub>H phage display library. Two phagemid transfected <em>E. coli</em> clones (P3-1 and P3-3) produced humanized-V<sub>H</sub>H, while another clone (P3-7) produced humanized-VH. At the optimal venom:antibody ratio, the VH/V<sub>H</sub>H purified from the <em>E. coli</em> homogenates neutralized PLA<sub>2</sub> enzyme activity comparable to the horse immune serum against the <em>N. kaouthia</em> holo-venom. Homology modeling and molecular docking revealed that the VH/V<sub>H</sub>H covered the areas around the PLA<sub>2</sub> catalytic groove and inserted their Complementarity Determining Regions (CDRs) into the enzymatic cleft. It is envisaged that the VH/V<sub>H</sub>H would ameliorate/abrogate the principal toxicity of the venom PLA<sub>2</sub> (membrane phospholipid catabolism leading to cellular and subcellular membrane damage which consequently causes hemolysis, hemorrhage, and dermo-/myo-necrosis), if they were used for passive immunotherapy of the cobra bitten victim. The speculation needs further investigations.http://www.mdpi.com/2072-6651/4/7/554snake bitesnake venomphospholipase A2 (PLA<sub>2</sub>)single domain antibody (SdAb)VH/V<sub>H</sub>Hhomology modelingmolecular docking |
| spellingShingle | Wanpen Chaicumpa Nitat Sookrung Kunan Bangphoomi Charnwit Chavanayarn Jeeraphong Thanongsaksrikul Kanyarat Thueng-in Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity Toxins snake bite snake venom phospholipase A2 (PLA<sub>2</sub>) single domain antibody (SdAb) VH/V<sub>H</sub>H homology modeling molecular docking |
| title | Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity |
| title_full | Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity |
| title_fullStr | Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity |
| title_full_unstemmed | Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity |
| title_short | Humanized-Single Domain Antibodies (VH/V<sub>H</sub>H) that Bound Specifically to<em> Naja kaouthia</em> Phospholipase A2 and Neutralized the Enzymatic Activity |
| title_sort | humanized single domain antibodies vh v lt sub gt h lt sub gt h that bound specifically to lt em gt naja kaouthia lt em gt phospholipase a2 and neutralized the enzymatic activity |
| topic | snake bite snake venom phospholipase A2 (PLA<sub>2</sub>) single domain antibody (SdAb) VH/V<sub>H</sub>H homology modeling molecular docking |
| url | http://www.mdpi.com/2072-6651/4/7/554 |
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