Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs
<b>Background</b>: Introducing unnatural base pairs into a natural, double-stranded DNA construct is a powerful tool within synthetic biology. Accordingly, the ability to detect these unnatural base pairs has many applications, including the study and detection of semisynthetic organisms...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-11-01
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| Series: | Biologics |
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| Online Access: | https://www.mdpi.com/2673-8449/4/4/26 |
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| author | Antonietta M. Lillo Nileena Velappan Ruilian Wu Madeline R. Bolding |
| author_facet | Antonietta M. Lillo Nileena Velappan Ruilian Wu Madeline R. Bolding |
| author_sort | Antonietta M. Lillo |
| collection | DOAJ |
| description | <b>Background</b>: Introducing unnatural base pairs into a natural, double-stranded DNA construct is a powerful tool within synthetic biology. Accordingly, the ability to detect these unnatural base pairs has many applications, including the study and detection of semisynthetic organisms. <b>Objective and Methods</b>: The work described here aimed to select human antibodies for the specific recognition of Hirao’s base pair dDs–dPn in various natural DNA contexts by using a combination of phage and yeast display technologies. <b>Results</b>: We selected a total of six antibodies in yeast-displayed scFv format, and further tested three of these antibodies in soluble form as minibodies and IgGs. We also describe an assay that can be used to detect plasmids containing dDs–dPn pair. <b>Conclusions</b>: Our antibodies did not afford the desired specificity or sensitivity for detection of a single unnatural base pair among thousands of natural. However, our data indicate not only that such detection is possible but also that these antibodies may be candidates for further affinity and specificity maturation. |
| format | Article |
| id | doaj-art-576bddc2dc464de3b6d8eca7f242ea07 |
| institution | DOAJ |
| issn | 2673-8449 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biologics |
| spelling | doaj-art-576bddc2dc464de3b6d8eca7f242ea072025-08-20T02:53:27ZengMDPI AGBiologics2673-84492024-11-014442344310.3390/biologics4040026Selection and Characterization of Antibodies Recognizing Unnatural Base PairsAntonietta M. Lillo0Nileena Velappan1Ruilian Wu2Madeline R. Bolding3Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USABioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USABioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USABioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA<b>Background</b>: Introducing unnatural base pairs into a natural, double-stranded DNA construct is a powerful tool within synthetic biology. Accordingly, the ability to detect these unnatural base pairs has many applications, including the study and detection of semisynthetic organisms. <b>Objective and Methods</b>: The work described here aimed to select human antibodies for the specific recognition of Hirao’s base pair dDs–dPn in various natural DNA contexts by using a combination of phage and yeast display technologies. <b>Results</b>: We selected a total of six antibodies in yeast-displayed scFv format, and further tested three of these antibodies in soluble form as minibodies and IgGs. We also describe an assay that can be used to detect plasmids containing dDs–dPn pair. <b>Conclusions</b>: Our antibodies did not afford the desired specificity or sensitivity for detection of a single unnatural base pair among thousands of natural. However, our data indicate not only that such detection is possible but also that these antibodies may be candidates for further affinity and specificity maturation.https://www.mdpi.com/2673-8449/4/4/26unnatural base pairsHirao’s base pairrecognition of unnatural base pairsphage displayyeast displayantibody selection |
| spellingShingle | Antonietta M. Lillo Nileena Velappan Ruilian Wu Madeline R. Bolding Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs Biologics unnatural base pairs Hirao’s base pair recognition of unnatural base pairs phage display yeast display antibody selection |
| title | Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs |
| title_full | Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs |
| title_fullStr | Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs |
| title_full_unstemmed | Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs |
| title_short | Selection and Characterization of Antibodies Recognizing Unnatural Base Pairs |
| title_sort | selection and characterization of antibodies recognizing unnatural base pairs |
| topic | unnatural base pairs Hirao’s base pair recognition of unnatural base pairs phage display yeast display antibody selection |
| url | https://www.mdpi.com/2673-8449/4/4/26 |
| work_keys_str_mv | AT antoniettamlillo selectionandcharacterizationofantibodiesrecognizingunnaturalbasepairs AT nileenavelappan selectionandcharacterizationofantibodiesrecognizingunnaturalbasepairs AT ruilianwu selectionandcharacterizationofantibodiesrecognizingunnaturalbasepairs AT madelinerbolding selectionandcharacterizationofantibodiesrecognizingunnaturalbasepairs |