Development of a Recombinant Monospecific Anti-PLGF Bivalent Nanobody and Evaluation of it in Angiogenesis Modulation
During the past two decades, tumor therapy based on monoclonal antibody has been found as a confident therapeutic approach in solid tumors and hematologic malignancies. Nanobodies are the smallest fragment of an antigen-binding domain in heavy chain-only antibody originated from the Camelidae family...
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| Veröffentlicht in: | Molecular biotechnology 2020-12, Vol.62 (11-12), p.580-588 |
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| creator | Nikooharf, Abolfazl Arezumand, Roghaye Mansouri, Kamran Khoshi, Amir Hossein Namdar Ahmadabad, Hassan |
| description | During the past two decades, tumor therapy based on monoclonal antibody has been found as a confident therapeutic approach in solid tumors and hematologic malignancies. Nanobodies are the smallest fragment of an antigen-binding domain in heavy chain-only antibody originated from the Camelidae family. Accordingly, they are being recently developed rapidly as diagnostic and therapeutic agents. In this regard, targeting of angiogenic factors like Placenta growth factor (PLGF) via nanobodies show a high effectiveness. In the current study, we developed a recombinant anti-PLGF bivalent nanobody based on the affinity enhancement mutant form of anti-PLGF nanobody to suppress the angiogenesis progression. Thereafter, the bivalent nanobody (bi-Nb) was cloned and then expressed into a bacterial system. Afterward, the purity was authorized using western blot assay and the affinity was assessed using ELISA. In this regard, proliferation, 3D capillary tube formation, and migration assays were employed as functional assays. The obtained data were analyzed using
t
-test and
P
|
| doi_str_mv | 10.1007/s12033-020-00275-7 |
| format | Article |
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t
-test and
P
< 0.05 was considered as statistically significant. The results indicate that the bivalent nanobody could inhibit proliferation, mobility, and formation of endothelial cell capillary-like structure. Moreover, the EC50 was estimated for endothelial cell’s proliferation and capillary tube’s formation to be about 100 ng/ml and 65 ng/ml, respectively. Migration of MCF-7 was inhibited as about 69%, rather than the control. Accumulation of data have shown that targeting of angiogenic factors like VEGF via monoclonal antibodies or nanobodies can be useful in the suppression of tumor progression. Also, the inhibition of PLGF with monoclonal antibody indicated that it is significant in angiogenesis suppression. However, due to intrinsic properties of nanobodies, they are suggested to be used. Since the small size is rapidly removed through liver or kidney system, so it is important to use bivalent or polymeric forms for extending the half-life. Our findings indicated that the inhibition of PLGF can prevent growth and proliferation of endothelial cells and tumor cells through the bivalent nanobody. So, it is suggested as a novel therapeutic agent for angiogenesis suppression.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>DOI: 10.1007/s12033-020-00275-7</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Biochemistry ; Biological Techniques ; Biotechnology ; Cell Biology ; Chemistry ; Chemistry and Materials Science ; Human Genetics ; Original Paper ; Protein Science</subject><ispartof>Molecular biotechnology, 2020-12, Vol.62 (11-12), p.580-588</ispartof><rights>Springer Science+Business Media, LLC, part of Springer Nature 2020</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c324t-1ffe9de3e2bfc5b908c6f42e472c6c5ba90a653d3d598e972e639d3d6e08382a3</citedby><cites>FETCH-LOGICAL-c324t-1ffe9de3e2bfc5b908c6f42e472c6c5ba90a653d3d598e972e639d3d6e08382a3</cites><orcidid>0000-0003-4197-2490</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12033-020-00275-7$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12033-020-00275-7$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids></links><search><creatorcontrib>Nikooharf, Abolfazl</creatorcontrib><creatorcontrib>Arezumand, Roghaye</creatorcontrib><creatorcontrib>Mansouri, Kamran</creatorcontrib><creatorcontrib>Khoshi, Amir Hossein</creatorcontrib><creatorcontrib>Namdar Ahmadabad, Hassan</creatorcontrib><title>Development of a Recombinant Monospecific Anti-PLGF Bivalent Nanobody and Evaluation of it in Angiogenesis Modulation</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><description>During the past two decades, tumor therapy based on monoclonal antibody has been found as a confident therapeutic approach in solid tumors and hematologic malignancies. Nanobodies are the smallest fragment of an antigen-binding domain in heavy chain-only antibody originated from the Camelidae family. Accordingly, they are being recently developed rapidly as diagnostic and therapeutic agents. In this regard, targeting of angiogenic factors like Placenta growth factor (PLGF) via nanobodies show a high effectiveness. In the current study, we developed a recombinant anti-PLGF bivalent nanobody based on the affinity enhancement mutant form of anti-PLGF nanobody to suppress the angiogenesis progression. Thereafter, the bivalent nanobody (bi-Nb) was cloned and then expressed into a bacterial system. Afterward, the purity was authorized using western blot assay and the affinity was assessed using ELISA. In this regard, proliferation, 3D capillary tube formation, and migration assays were employed as functional assays. The obtained data were analyzed using
t
-test and
P
< 0.05 was considered as statistically significant. The results indicate that the bivalent nanobody could inhibit proliferation, mobility, and formation of endothelial cell capillary-like structure. Moreover, the EC50 was estimated for endothelial cell’s proliferation and capillary tube’s formation to be about 100 ng/ml and 65 ng/ml, respectively. Migration of MCF-7 was inhibited as about 69%, rather than the control. Accumulation of data have shown that targeting of angiogenic factors like VEGF via monoclonal antibodies or nanobodies can be useful in the suppression of tumor progression. Also, the inhibition of PLGF with monoclonal antibody indicated that it is significant in angiogenesis suppression. However, due to intrinsic properties of nanobodies, they are suggested to be used. Since the small size is rapidly removed through liver or kidney system, so it is important to use bivalent or polymeric forms for extending the half-life. Our findings indicated that the inhibition of PLGF can prevent growth and proliferation of endothelial cells and tumor cells through the bivalent nanobody. So, it is suggested as a novel therapeutic agent for angiogenesis suppression.</description><subject>Biochemistry</subject><subject>Biological Techniques</subject><subject>Biotechnology</subject><subject>Cell Biology</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Human Genetics</subject><subject>Original Paper</subject><subject>Protein Science</subject><issn>1073-6085</issn><issn>1559-0305</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNp9kDtPwzAYRSMEEqXwB5gyshj8iJ1kLKUtSOUhBLPlOJ8rV4ld4qRS_z1uw8xk--qcK_kmyS3B9wTj_CEQihlDmGKEMc05ys-SCeG8RJhhfh7vOGdI4IJfJlchbCNEeMYmyfAEe2j8rgXXp96kKv0E7dvKOhWDV-982IG2xup05nqLPtarZfpo96o5Cm_K-crXh1S5Ol3EcFC99e5YZPvUuuhsrN-Ag2BDbKuH5gRcJxdGNQFu_s5p8r1cfM2f0fp99TKfrZFmNOsRMQbKGhjQymhelbjQwmQUspxqEQNVYiU4q1nNywLKnIJgZXwJwAUrqGLT5G7s3XX-Z4DQy9YGDU2jHPghSJplQohScBJROqK68yF0YOSus63qDpJgedxYjhvLuLE8bSzzKLFRChF2G-jk1g-di1_6z_oFvk2AUw</recordid><startdate>20201201</startdate><enddate>20201201</enddate><creator>Nikooharf, Abolfazl</creator><creator>Arezumand, Roghaye</creator><creator>Mansouri, Kamran</creator><creator>Khoshi, Amir Hossein</creator><creator>Namdar Ahmadabad, Hassan</creator><general>Springer US</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-4197-2490</orcidid></search><sort><creationdate>20201201</creationdate><title>Development of a Recombinant Monospecific Anti-PLGF Bivalent Nanobody and Evaluation of it in Angiogenesis Modulation</title><author>Nikooharf, Abolfazl ; Arezumand, Roghaye ; Mansouri, Kamran ; Khoshi, Amir Hossein ; Namdar Ahmadabad, Hassan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c324t-1ffe9de3e2bfc5b908c6f42e472c6c5ba90a653d3d598e972e639d3d6e08382a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Biochemistry</topic><topic>Biological Techniques</topic><topic>Biotechnology</topic><topic>Cell Biology</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Human Genetics</topic><topic>Original Paper</topic><topic>Protein Science</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nikooharf, Abolfazl</creatorcontrib><creatorcontrib>Arezumand, Roghaye</creatorcontrib><creatorcontrib>Mansouri, Kamran</creatorcontrib><creatorcontrib>Khoshi, Amir Hossein</creatorcontrib><creatorcontrib>Namdar Ahmadabad, Hassan</creatorcontrib><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nikooharf, Abolfazl</au><au>Arezumand, Roghaye</au><au>Mansouri, Kamran</au><au>Khoshi, Amir Hossein</au><au>Namdar Ahmadabad, Hassan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a Recombinant Monospecific Anti-PLGF Bivalent Nanobody and Evaluation of it in Angiogenesis Modulation</atitle><jtitle>Molecular biotechnology</jtitle><stitle>Mol Biotechnol</stitle><date>2020-12-01</date><risdate>2020</risdate><volume>62</volume><issue>11-12</issue><spage>580</spage><epage>588</epage><pages>580-588</pages><issn>1073-6085</issn><eissn>1559-0305</eissn><abstract>During the past two decades, tumor therapy based on monoclonal antibody has been found as a confident therapeutic approach in solid tumors and hematologic malignancies. Nanobodies are the smallest fragment of an antigen-binding domain in heavy chain-only antibody originated from the Camelidae family. Accordingly, they are being recently developed rapidly as diagnostic and therapeutic agents. In this regard, targeting of angiogenic factors like Placenta growth factor (PLGF) via nanobodies show a high effectiveness. In the current study, we developed a recombinant anti-PLGF bivalent nanobody based on the affinity enhancement mutant form of anti-PLGF nanobody to suppress the angiogenesis progression. Thereafter, the bivalent nanobody (bi-Nb) was cloned and then expressed into a bacterial system. Afterward, the purity was authorized using western blot assay and the affinity was assessed using ELISA. In this regard, proliferation, 3D capillary tube formation, and migration assays were employed as functional assays. The obtained data were analyzed using
t
-test and
P
< 0.05 was considered as statistically significant. The results indicate that the bivalent nanobody could inhibit proliferation, mobility, and formation of endothelial cell capillary-like structure. Moreover, the EC50 was estimated for endothelial cell’s proliferation and capillary tube’s formation to be about 100 ng/ml and 65 ng/ml, respectively. Migration of MCF-7 was inhibited as about 69%, rather than the control. Accumulation of data have shown that targeting of angiogenic factors like VEGF via monoclonal antibodies or nanobodies can be useful in the suppression of tumor progression. Also, the inhibition of PLGF with monoclonal antibody indicated that it is significant in angiogenesis suppression. However, due to intrinsic properties of nanobodies, they are suggested to be used. Since the small size is rapidly removed through liver or kidney system, so it is important to use bivalent or polymeric forms for extending the half-life. Our findings indicated that the inhibition of PLGF can prevent growth and proliferation of endothelial cells and tumor cells through the bivalent nanobody. So, it is suggested as a novel therapeutic agent for angiogenesis suppression.</abstract><cop>New York</cop><pub>Springer US</pub><doi>10.1007/s12033-020-00275-7</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0003-4197-2490</orcidid></addata></record> |
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| subjects | Biochemistry Biological Techniques Biotechnology Cell Biology Chemistry Chemistry and Materials Science Human Genetics Original Paper Protein Science |
| title | Development of a Recombinant Monospecific Anti-PLGF Bivalent Nanobody and Evaluation of it in Angiogenesis Modulation |
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