A Switchable Site-Specific Antibody Conjugate

Lyu, Zhigang; Liu, Kang; Buuh, Zakey Yusuf; Jiang, Dawei; Du, Juanjuan; Wissler, Haley L.; Cai, Weibo; Wang, Rongsheng E.

doi:10.1021/acschembio.8b00107

Cited by 14 publications

(19 citation statements)

References 31 publications

(85 reference statements)

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“…With DUPA conjugating to pAcF site-specifically expressed on anti-CD3 Fab, the EC 50 of the conjugate has been reduced to 100pM, and serum half-life has been improved to 5–6 h, resulting effective in a treatment in prostate cancer xenograft mouse models [ 81 ]. Same strategy was also employed to synthesize an anti-CD3 Fab-folate conjugate [ 82 ] and a switchable αGCN4-Fab conjugate [ 83 ].…”

Section: Bispecific Antibody (Bsab)mentioning

confidence: 99%

Therapeutic applications of genetic code expansion

Huang

Liu

2018

Synthetic and Systems Biotechnology

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In nature, a limited, conservative set of amino acids are utilized to synthesize proteins. Genetic code expansion technique reassigns codons and incorporates noncanonical amino acids (ncAAs) through orthogonal aminoacyl-tRNA synthetase (aaRS)/tRNA pairs. The past decade has witnessed the rapid growth in diversity and scope for therapeutic applications of this technology. Here, we provided an update on the recent progress using genetic code expansion in the following areas: antibody-drug conjugates (ADCs), bispecific antibodies (BsAb), immunotherapies, long-lasting protein therapeutics, biosynthesized peptides, engineered viruses and cells, as well as other therapeutic related applications, where the technique was used to elucidate the mechanisms of biotherapeutics and drug targets.

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Section: Bispecific Antibody (Bsab)mentioning

confidence: 99%

Therapeutic applications of genetic code expansion

Huang

Liu

2018

Synthetic and Systems Biotechnology

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“…51 The plasmid (pBAD_αPD-L1) was constructed to harbor the heavy-and lambda light chain genes of the Fab fragment following a stll signal peptide. 36,52 The wild type αPD-L1 Fab was expressed in Escherichia coli, with a yield of 4.2 mg/L in shake flask cultures. Sodium dodecyl sulfate/ polyacrylamide gel electrophoresis (SDS/PAGE) analysis showed that the Fab fragment migrated as a single band, with >95% purity and a molecular mass of ∼45 kDa (Figure S2).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…33,34 In this approach, the amber stop codon employs paired orthogonal amino-acyl transferase and iso-tRNA to incorporate the desired UAA site specifically into proteins including the recombinant antibody. 33,34 Antibodies incorporating UAAs, such as pacetylphenylalanine (pAcF), can be coupled to small molecules of interest via a stable linkage, 29,35,36 generating site-specific therapeutics in high yields. The UAA conjugates as therapeutic candidates have demonstrated superior in vivo efficacy, stability, and toxicology profiles to the cysteine conjugates.…”

Section: ■ Introductionmentioning

confidence: 99%

Site-Specific Immuno-PET Tracer to Image PD-L1

et al. 2019

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The rapid ascension of immune checkpoint blockade treatments has placed an emphasis on the need for viable, robust, and noninvasive imaging methods for immune checkpoint proteins, which could be of diagnostic value. Immunoconjugate-based positron emission tomography (immuno-PET) allows for sensitive and quantitative imaging of target levels and has promising potential for the noninvasive evaluation of immune checkpoint proteins. However, the advancement of immuno-PET is currently limited by available imaging tools, which heavily rely on full-length IgGs with Fc-mediated effects and are heterogeneous mixtures upon random conjugation with chelators for imaging. Herein, we have developed a site-specific αPD-L1 Fab conjugate with the chelator 1,4,7triazacyclononane-N,N′,N″-triacetic acid (NOTA), enabling radiolabeling for PET imaging, using the amber suppression-mediated genetic incorporation of unnatural amino acid (UAA), pazidophenylalanine. This Fab conjugate is homogeneous and demonstrated tight binding toward the PD-L1 antigen in vitro. The radiolabeled version, 64 Cu-NOTA-αPD-L1, has been employed in PET imaging to allow for effective visualization and mapping of the biodistribution of PD-L1 in two normal mouse models, including the capturing of different PD-L1 expression levels in the spleens of the different mouse types. Follow-up in vivo blocking studies and ex vivo fluorescent staining further validated specific tissue uptakes of the imaging agent. This approach illustrates the utility of UAA-based site-specific Fab conjugation as a general strategy for making sensitive PET imaging probes, which could facilitate the elucidation of the roles of a wide variety of immune checkpoint proteins in immunotherapy.

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“…Currently there are 2 strategies: site-specific conjugation technology and chemical structural modification of the linker‒antibody attachment. Site-specific conjugation through antibody engineering is extensively reviewed elsewhere 60 , 61 , 62 .…”

Section: Linker‒antibody Attachments Of the Linkermentioning

confidence: 99%

Antibody–drug conjugates: Recent advances in linker chemistry

Xiao²,

Xie³

et al. 2021

Acta Pharmaceutica Sinica B

172

114

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Antibody–drug conjugates (ADCs) are gradually revolutionizing clinical cancer therapy. The antibody–drug conjugate linker molecule determines both the efficacy and the adverse effects, and so has a major influence on the fate of ADCs. An ideal linker should be stable in the circulatory system and release the cytotoxic payload specifically in the tumor. However, existing linkers often release payloads nonspecifically and inevitably lead to off-target toxicity. This defect is becoming an increasingly important factor that restricts the development of ADCs. The pursuit of ADCs with optimal therapeutic windows has resulted in remarkable progress in the discovery and development of novel linkers. The present review summarizes the advance of the chemical trigger, linker‒antibody attachment and linker‒payload attachment over the last 5 years, and describes the ADMET properties of ADCs. This work also helps clarify future developmental directions for the linkers.

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A Switchable Site-Specific Antibody Conjugate

Cited by 14 publications

References 31 publications

Therapeutic applications of genetic code expansion

Therapeutic applications of genetic code expansion

Site-Specific Immuno-PET Tracer to Image PD-L1

Antibody–drug conjugates: Recent advances in linker chemistry

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