A unique algorithm for the determination of peptide-carrier protein conjugation ratio by amino acid analysis using intrinsic internal standard
•A new algorithm to calculate peptide-carrier protein conjugate ratio•An intrinsic amino acid application to minimize the assay variability•The optimized method was applied to various FP-rTTHc conjugate candidates An N-terminal peptide of the HIV-1 fusion peptide (FP) with eight amino acid residues...
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| Veröffentlicht in: | Vaccine 2020-06, Vol.38 (29), p.4507-4511 |
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| container_issue | 29 |
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| container_title | Vaccine |
| container_volume | 38 |
| creator | Yang, Gengcheng J. Yang, Yanhong Shaddeau, Andrew Cai, Cindy X. Li, Yile Gulla, Krishana Zhang, Yaqiu Ou, Li Cooper, Jonathan W. Lei, Q. Paula |
| description | •A new algorithm to calculate peptide-carrier protein conjugate ratio•An intrinsic amino acid application to minimize the assay variability•The optimized method was applied to various FP-rTTHc conjugate candidates
An N-terminal peptide of the HIV-1 fusion peptide (FP) with eight amino acid residues (FP8) was conjugated to a recombinant Tetanus Toxoid Heavy Chain Fragment C (rTTHc) as a carrier protein to help boosting immunogenicity against HIV-1. In this rapid communication, a unique algorithm to determine FP-rTTHc conjugation ratio was developed based off the amino acid analysis. Five well recovered amino acids (present in both FP and rTTHc) were used to calculate the conjugation ratio, while proline (present only in rTTHc) was identified and utilized as the intrinsic internal standard for normalization. With this calculation, the assay variability was minimized ( |
| doi_str_mv | 10.1016/j.vaccine.2020.04.080 |
| format | Article |
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An N-terminal peptide of the HIV-1 fusion peptide (FP) with eight amino acid residues (FP8) was conjugated to a recombinant Tetanus Toxoid Heavy Chain Fragment C (rTTHc) as a carrier protein to help boosting immunogenicity against HIV-1. In this rapid communication, a unique algorithm to determine FP-rTTHc conjugation ratio was developed based off the amino acid analysis. Five well recovered amino acids (present in both FP and rTTHc) were used to calculate the conjugation ratio, while proline (present only in rTTHc) was identified and utilized as the intrinsic internal standard for normalization. With this calculation, the assay variability was minimized (<20%), especially for conjugates with moderate to low conjugation ratios as being compared to previously reported methods. The approach offers a reliable tool to determine the efficiency of the conjugation reactions for in-process monitoring and for final conjugate product characterization.</description><identifier>ISSN: 0264-410X</identifier><identifier>EISSN: 1873-2518</identifier><identifier>DOI: 10.1016/j.vaccine.2020.04.080</identifier><identifier>PMID: 32448620</identifier><language>eng</language><publisher>Netherlands: Elsevier Ltd</publisher><subject>Algorithms ; Amino acid analysis ; Amino Acids ; Antigens ; Carrier protein ; Carrier Proteins ; Conjugates ; Conjugation ; Conjugation ratio ; Fusion peptide ; HIV ; HIV-1 vaccine ; Human immunodeficiency virus ; Immunogenicity ; Mathematical analysis ; Methods ; Peptides ; Proline ; Proteins ; Reference Standards ; rTTHc ; Tetanus ; Vaccines</subject><ispartof>Vaccine, 2020-06, Vol.38 (29), p.4507-4511</ispartof><rights>2020</rights><rights>Published by Elsevier Ltd.</rights><rights>Copyright Elsevier Limited Jun 15, 2020</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c393t-6240c7ca9c15ec6b91b4040e5b12c20dd0a91bae1dc6a613d860fff71164aa4d3</citedby><cites>FETCH-LOGICAL-c393t-6240c7ca9c15ec6b91b4040e5b12c20dd0a91bae1dc6a613d860fff71164aa4d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.proquest.com/docview/2425692020?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993,64383,64385,64387,72239</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32448620$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yang, Gengcheng J.</creatorcontrib><creatorcontrib>Yang, Yanhong</creatorcontrib><creatorcontrib>Shaddeau, Andrew</creatorcontrib><creatorcontrib>Cai, Cindy X.</creatorcontrib><creatorcontrib>Li, Yile</creatorcontrib><creatorcontrib>Gulla, Krishana</creatorcontrib><creatorcontrib>Zhang, Yaqiu</creatorcontrib><creatorcontrib>Ou, Li</creatorcontrib><creatorcontrib>Cooper, Jonathan W.</creatorcontrib><creatorcontrib>Lei, Q. Paula</creatorcontrib><title>A unique algorithm for the determination of peptide-carrier protein conjugation ratio by amino acid analysis using intrinsic internal standard</title><title>Vaccine</title><addtitle>Vaccine</addtitle><description>•A new algorithm to calculate peptide-carrier protein conjugate ratio•An intrinsic amino acid application to minimize the assay variability•The optimized method was applied to various FP-rTTHc conjugate candidates
An N-terminal peptide of the HIV-1 fusion peptide (FP) with eight amino acid residues (FP8) was conjugated to a recombinant Tetanus Toxoid Heavy Chain Fragment C (rTTHc) as a carrier protein to help boosting immunogenicity against HIV-1. In this rapid communication, a unique algorithm to determine FP-rTTHc conjugation ratio was developed based off the amino acid analysis. Five well recovered amino acids (present in both FP and rTTHc) were used to calculate the conjugation ratio, while proline (present only in rTTHc) was identified and utilized as the intrinsic internal standard for normalization. With this calculation, the assay variability was minimized (<20%), especially for conjugates with moderate to low conjugation ratios as being compared to previously reported methods. The approach offers a reliable tool to determine the efficiency of the conjugation reactions for in-process monitoring and for final conjugate product characterization.</description><subject>Algorithms</subject><subject>Amino acid analysis</subject><subject>Amino Acids</subject><subject>Antigens</subject><subject>Carrier protein</subject><subject>Carrier Proteins</subject><subject>Conjugates</subject><subject>Conjugation</subject><subject>Conjugation ratio</subject><subject>Fusion peptide</subject><subject>HIV</subject><subject>HIV-1 vaccine</subject><subject>Human immunodeficiency virus</subject><subject>Immunogenicity</subject><subject>Mathematical analysis</subject><subject>Methods</subject><subject>Peptides</subject><subject>Proline</subject><subject>Proteins</subject><subject>Reference Standards</subject><subject>rTTHc</subject><subject>Tetanus</subject><subject>Vaccines</subject><issn>0264-410X</issn><issn>1873-2518</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkU2O1DAQhS0EYpqBI4AssWGTUHYcd7JCo9HwI43EBiR2lmNXehwldmM7I_UlODOOumHBhlVZ9veqXO8R8ppBzYDJ91P9qI1xHmsOHGoQNXTwhOxYt28q3rLuKdkBl6ISDH5ckRcpTQDQNqx_Tq4aLkQnOezIrxu6evdzRarnQ4guPyx0DJHmB6QWM8bFeZ1d8DSM9IjH7CxWRsfoMNJjDBmdpyb4aT2csbgVOpyoLspAtXGWaq_nU3KJrsn5A3U-R-eTM9sJY3mkKWtvdbQvybNRzwlfXeo1-f7x7tvt5-r-66cvtzf3lWn6JleSCzB7o3vDWjRy6NkgQAC2A-OGg7Wgy5VGZo3UkjW2kzCO454xKbQWtrkm7859ywpl-ZTV4pLBedYew5pU6S_bfcPbrqBv_0GnsG6f3ijeyn7zv1DtmTIxpBRxVMfoFh1PioHaAlOTugSmNoECoUpgRffm0n0dFrR_VX8SKsCHM4DFjsfiukrGoTdoXUSTlQ3uPyN-A9GarJg</recordid><startdate>20200615</startdate><enddate>20200615</enddate><creator>Yang, Gengcheng J.</creator><creator>Yang, Yanhong</creator><creator>Shaddeau, Andrew</creator><creator>Cai, Cindy X.</creator><creator>Li, Yile</creator><creator>Gulla, Krishana</creator><creator>Zhang, Yaqiu</creator><creator>Ou, Li</creator><creator>Cooper, Jonathan W.</creator><creator>Lei, Q. 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Paula</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A unique algorithm for the determination of peptide-carrier protein conjugation ratio by amino acid analysis using intrinsic internal standard</atitle><jtitle>Vaccine</jtitle><addtitle>Vaccine</addtitle><date>2020-06-15</date><risdate>2020</risdate><volume>38</volume><issue>29</issue><spage>4507</spage><epage>4511</epage><pages>4507-4511</pages><issn>0264-410X</issn><eissn>1873-2518</eissn><abstract>•A new algorithm to calculate peptide-carrier protein conjugate ratio•An intrinsic amino acid application to minimize the assay variability•The optimized method was applied to various FP-rTTHc conjugate candidates
An N-terminal peptide of the HIV-1 fusion peptide (FP) with eight amino acid residues (FP8) was conjugated to a recombinant Tetanus Toxoid Heavy Chain Fragment C (rTTHc) as a carrier protein to help boosting immunogenicity against HIV-1. In this rapid communication, a unique algorithm to determine FP-rTTHc conjugation ratio was developed based off the amino acid analysis. Five well recovered amino acids (present in both FP and rTTHc) were used to calculate the conjugation ratio, while proline (present only in rTTHc) was identified and utilized as the intrinsic internal standard for normalization. With this calculation, the assay variability was minimized (<20%), especially for conjugates with moderate to low conjugation ratios as being compared to previously reported methods. The approach offers a reliable tool to determine the efficiency of the conjugation reactions for in-process monitoring and for final conjugate product characterization.</abstract><cop>Netherlands</cop><pub>Elsevier Ltd</pub><pmid>32448620</pmid><doi>10.1016/j.vaccine.2020.04.080</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0264-410X |
| ispartof | Vaccine, 2020-06, Vol.38 (29), p.4507-4511 |
| issn | 0264-410X 1873-2518 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2406573258 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present); ProQuest Central UK/Ireland |
| subjects | Algorithms Amino acid analysis Amino Acids Antigens Carrier protein Carrier Proteins Conjugates Conjugation Conjugation ratio Fusion peptide HIV HIV-1 vaccine Human immunodeficiency virus Immunogenicity Mathematical analysis Methods Peptides Proline Proteins Reference Standards rTTHc Tetanus Vaccines |
| title | A unique algorithm for the determination of peptide-carrier protein conjugation ratio by amino acid analysis using intrinsic internal standard |
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