Harnessing a catalytic lysine residue for the one-step preparation of homogeneous antibody-drug conjugates

Nanna, Alex R.; Li, Xiuling; Walseng, Even; Pedzisa, Lee; Goydel, Rebecca S.; Hymel, David; Burke, Terrence R.; Roush, William; Rader, Christoph

doi:10.1038/s41467-017-01257-1

Cited by 79 publications

(127 citation statements)

References 70 publications

Supporting

Mentioning

125

Contrasting

Order By: Relevance

“…Novel conjugation methods which are more specific and potent in conjugating drug payloads to antibodies are also evolving. 38 More interestingly, non-internalising ADC that releases its drugs upon a click reaction with a chemical activator was recently developed, which showed efficacy in treating cancer that are resistant to those ADCs must be internalized. 39 In summary, this study reports a neo-antigen epitope generated by abnromal post-translational modification of glycoproteins, with high positivity in a broad spectrum of cancer types.…”

Section: Discussionmentioning

confidence: 99%

An antibody drug conjugate targeting a GSTA glycosite-signature epitope of mucin1 expressed by non-small cell lung cancer

Pan

Tang

Jiao

et al. 2019

Preprint

View full text Add to dashboard Cite

Background: Antibodies targeting abnormally glycosylated proteins have been ineffective in treating cancer. Antibody-drug conjugates are emerging as an efficient option, which allow specific delivery of drugs into tumors. We and others have dissected the abnormally glycosylated tandem repeat region of MUC1 glycoprotein as three site-specific glycosylated neoantigen peptide motifs (PDTR, GSTA, GVTS) for monoclonal antibody binding. Methods:of monoclonal antibodies was studied by immunofluorescence staining and colocalization with lysosomal markers in live cells. Antibody positivity in tumor and peritumoral tissue samples were studied by immunohistochemistry. The efficacy of anti-MUC1 ADCs were evaluated with various cancer cell lines and mouse tumor xenograft model. Results: We describe an anti-MUC1 ADC by conjugating GSTA neoantigen-specific 16A with monomethyl auristatin E (MMAE). 16A-MMAE showed potent antitumoral efficacy with IC 50 ranging from 0.2 to 49.4 nM toward multiple types of cancer cells. In vivo, 16A-MMAE showed dose-dependent inhibition of tumor growth in mouse xenograft of NCI-H838 NSCLC cell line, with minimum effective dose at 1 mg/kg. At the dose of 3 mg/kg, 16A-MMAE did not cause significant toxicity in a transgenic mouse expressing human MUC1. Conclusions: The high antitumoral efficacy of 16A-MMAE suggest that aberrant glycosylated MUC1 neoantigen is a target with high positivity in multiple cancer types for ADC development. Personalized therapy may be achieved by development of glycosite-specific antibody-drug conjugates.

show abstract

Section: Discussionmentioning

confidence: 99%

An antibody drug conjugate targeting a GSTA glycosite-signature epitope of mucin1 expressed by non-small cell lung cancer

Pan

Tang

Jiao

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…The high reactivity of the electrophiles and abundance of nucleophilic residues with similar reactivity causes the reaction sites on the protein to act similarly, resulting in the heterogeneous conjugation. Tuning the reactivity of both the reagents and the microenvironment of a nucleophilic residue can improve the selectivity of the reactions . Alternatively, methods for site‐specific conjugation can provide full control of the reaction sites and number of modifications by introducing selectively modifiable handles.…”

Section: Introductionmentioning

confidence: 99%

Considerations on Probe Design for Affinity‐Guided Protein Conjugation

2019

View full text Add to dashboard Cite

The plethora of methods developed for the creation of protein conjugates often differs significantly with regard to the heterogeneity of the resulting products, in the degree of genetic manipulation of the protein required, and in the technical skills required to perform the conjugation procedure. Affinity‐guided protein conjugation is a protein labeling methodology based on noncovalent binding interactions between a labeling probe and the protein of interest. These interactions increase the local concentration of a reactive group in the probe on the protein surface thus facilitating the conjugation in proximity of the complexation site. The ability to produce high‐quality conjugates from nongenetically modified proteins both in vitro, but also in cells, demonstrates the power of affinity‐guided protein conjugation. Here, we present the progress of affinity‐guided protein conjugation in relation to selective protein labeling in living systems and the formation of high‐quality protein conjugates. Furthermore, the probe design will be discussed in relation to the utility of the probe for labeling in vitro or in living systems.

show abstract

“…On the other hand, protein engineering or other technologies can produce a more homogenous product by attaching drugs to specific conjugation sites. [18][19][20][21] For example, when engineered cysteine residues were introduced in the mAb using the THIOMAB™ platform technology, the final product, THIOMAB™ drug conjugate (TDC), showed a homogenous drug-to-antibody ratio (DAR) of 2, 22 whereas a conventional (interchain-cysteine) ADC has an average DAR of 3.5. Structures of a TDC with engineered cysteine in the heavy chain (HC) and an example of an interchain-cysteine ADC are shown in Figure 1.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Ring-Opening Reaction of Succinimide Linkers in ADCs

Zheng¹,

Chen²,

Wang³

et al. 2019

Journal of Pharmaceutical Sciences

View full text Add to dashboard Cite

A new class of highly potent biopharmaceutical drugs, antibody-drug conjugates (ADCs), has been proven to be clinically effective to treat oncologic diseases. ADCs contain 3 major components: the monoclonal antibody, cytotoxic drug, and chemical linker. THIOMAB™ drug conjugates and interchaincysteine ADCs are common ADC platforms that apply thiol-maleimide chemistry via Michael addition to conjugate linker-drugs to cysteine residues. However, the resulting succinimide ring in the linker is susceptible to ring-opening reactions via hydrolysis, especially at high pH and elevated temperatures. Once the succinimide ring is opened, in vivo stability of the ADCs can be changed and the therapeutic activity will be altered. In this study, we investigated the impact of conjugation sites on succinimide ring opening for ADCs. A new methodology based on imaged capillary isoelectric focusing was developed to monitor the formation of succinimide ring-opened products. In addition, a reversephase high-performance liquid chromatography method was used to monitor site-specific ringopening reactions. Our data confirmed that succinimide ring-opening rates in ADCs are conjugationsite dependent. With a good understanding of the conjugation site impact on final product's stability, it is potentially feasible to modify ring-opening rates in vitro to achieve desirable in vivo stability and biological activity.

show abstract

Harnessing a catalytic lysine residue for the one-step preparation of homogeneous antibody-drug conjugates

Cited by 79 publications

References 70 publications

An antibody drug conjugate targeting a GSTA glycosite-signature epitope of mucin1 expressed by non-small cell lung cancer

An antibody drug conjugate targeting a GSTA glycosite-signature epitope of mucin1 expressed by non-small cell lung cancer

Considerations on Probe Design for Affinity‐Guided Protein Conjugation

Characterization of Ring-Opening Reaction of Succinimide Linkers in ADCs

Contact Info

Product

Resources

About