Epitope mapping of anti-drug antibodies to a clinical candidate bispecific antibody

Schick, Arthur J.; Lundin, Victor; Low, Justin; Peng, Kun; Vandlen, Richard; Wecksler, Aaron T.

doi:10.1080/19420862.2022.2028337

Cited by 11 publications

(10 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 104 A recent proof-of-concept work by Genentech involves FPOP-based ADA epitope mapping of a clinical candidate bispecific antibody. 76 As reported, the CDRs of both heavy and light chains in both arms could be recognized by ADAs as per the FPOP-MS data. Interestingly, two additional regions on both arms were incrementally exposed in the presence of ADAs, suggesting the possibility of ADAs serving as allosteric modulators of TPs.…”

Section: Scope Of Bce Mapping Methodologiessupporting

confidence: 56%

See 1 more Smart Citation

T and B cell epitope analysis for the immunogenicity evaluation and mitigation of antibody-based therapeutics

Sun,

Qian,

Zhang

2024

mAbs

View full text Add to dashboard Cite

The surge in the clinical use of therapeutic antibodies has reshaped the landscape of pharmaceutical therapy for many diseases, including rare and challenging conditions. However, the administration of exogenous biologics could potentially trigger unwanted immune responses such as generation of anti-drug antibodies (ADAs). Real-world experiences have illuminated the clear correlation between the ADA occurrence and unsatisfactory therapeutic outcomes as well as immune-related adverse events. By retrospectively examining research involving immunogenicity analysis, we noticed the growing emphasis on elucidating the immunogenic epitope profiles of antibody-based therapeutics aiming for mechanistic understanding the immunogenicity generation and, ideally, mitigating the risks. As such, we have comprehensively summarized here the progress in both experimental and computational methodologies for the characterization of T and B cell epitopes of therapeutics. Furthermore, the successful practice of epitope-driven deimmunization of biotherapeutics is exceptionally highlighted in this article.

show abstract

Section: Scope Of Bce Mapping Methodologiessupporting

confidence: 56%

“…Interestingly, two additional regions on both arms were incrementally exposed in the presence of ADAs, suggesting the possibility of ADAs serving as allosteric modulators of TPs. 76 …”

Section: Scope Of Bce Mapping Methodologiesmentioning

confidence: 99%

T and B cell epitope analysis for the immunogenicity evaluation and mitigation of antibody-based therapeutics

Sun,

Qian,

Zhang

2024

mAbs

View full text Add to dashboard Cite

show abstract

“…The current DDA method incorporated a long exclusion time in addition to 6 technical replicates for all samples in order to increase the depth of the analysis. Compared to the literature, most studies using FPOP focus on a single non-oxidation control and biological triplicates [35,45]. Alternatively biological duplicates for both oxidized and non-oxidized control have been used [23,46].…”

Section: Discussionmentioning

confidence: 99%

Epitope mapping of SARS-CoV-2 RBDs by hydroxyl radical protein footprinting reveals the importance of including negative antibody controls

Larsen,

Kaczmarek,

Palarasah

et al. 2023

Preprint

View full text Add to dashboard Cite

Epitope mapping methods using hydroxyl radical protein footprinting (HRPF) are currently hampered by high initial cost and complex experimental setup. Here we set out to present a generally applicable method using Fenton chemistry for mapping of epitopes and paratopes in a standard laboratory setting. Furthermore, the described method illustrates the importance of controls on several levels when performing mass spectrometry-based epitope mapping. In particular, the inclusion of negative antibody controls has not previously been widely utilized in epitope mapping by HRPF analysis. In order to limit the false positives, we further introduced quantification by TMT labelling, thereby allowing for direct comparison between sample conditions and biological triplicates. Lastly, up to six technical replicates were incorporated in the experimental setup in order to achieve increased depth of the final analysis.Both binding and openings of regions on receptor binding domain (RBD) from SARS-CoV-2 Spike Protein, Alpha and Delta variants, were observed. The negative control antibody experiment, combined with the high overlap between biological triplicates, resulted in the exclusion of 40% of the significantly changed regions, including both binding and opening regions. The identified binding region agrees with the literature for neutralizing antibodies towards SARS-CoV-2 Spike Protein.The presented method is an easily implementable technique for the analysis of HRPF in a generic MS-based laboratory. The high reliability of the data was achieved by increasing the number of technical and biological replicates combined with negative antibody controls.

show abstract

“… Information obtained from HRPF: ( A ) epitope mapping, as in [ 20 ]; ( B ) allosteric changes induced by antigen binding [ 24 ]; ( C ) host response, as in [ 23 ], in which anti-drug antibodies exhibit protection in response to therapy; ( D ) aggregation interfaces, as in [ 22 ]. …”

Section: Figurementioning

confidence: 99%

“…Sharp et al showed the effective determination of the adalimumab epitope in trimeric TNFα using the commercially available flash oxidation (FOX) protein footprinting system [ 20 ]. Schick et al used HRPF to probe the primary epitopes for polyclonal anti-drug antibodies raised against a bi-specific mAb in a monkey pharmacological model [ 23 ], demonstrating the utility of hydroxyl radical footprinting not only in drug–target interactions but also in host drug response. Schoof et al used HRPF in conjunction with crystallography and cryo-EM to delineate an ultrapotent nanobody interaction with the SARS-CoV2 spike protein [ 24 ], while Sevillano et al used the method to improve the affinity maturation of a recombinant Fab inhibitor [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

Structural Investigation of Therapeutic Antibodies Using Hydroxyl Radical Protein Footprinting Methods

Ralston

Sharp

2022

Antibodies

View full text Add to dashboard Cite

Commercial monoclonal antibodies are growing and important components of modern therapies against a multitude of human diseases. Well-known high-resolution structural methods such as protein crystallography are often used to characterize antibody structures and to determine paratope and/or epitope binding regions in order to refine antibody design. However, many standard structural techniques require specialized sample preparation that may perturb antibody structure or require high concentrations or other conditions that are far from the conditions conducive to the accurate determination of antigen binding or kinetics. We describe here in this minireview the relatively new method of hydroxyl radical protein footprinting, a solution-state method that can provide structural and kinetic information on antibodies or antibody–antigen interactions useful for therapeutic antibody design. We provide a brief history of hydroxyl radical footprinting, examples of current implementations, and recent advances in throughput and accessibility.

show abstract

Epitope mapping of anti-drug antibodies to a clinical candidate bispecific antibody

Cited by 11 publications

References 25 publications

T and B cell epitope analysis for the immunogenicity evaluation and mitigation of antibody-based therapeutics

T and B cell epitope analysis for the immunogenicity evaluation and mitigation of antibody-based therapeutics

Epitope mapping of SARS-CoV-2 RBDs by hydroxyl radical protein footprinting reveals the importance of including negative antibody controls

Structural Investigation of Therapeutic Antibodies Using Hydroxyl Radical Protein Footprinting Methods

Contact Info

Product

Resources

About