Proximity-Induced Site-Specific Antibody Conjugation

Yu, Chenfei; Tang, Juan; Loredo, Axel; Chen, Yuda; Jung, Sung Yun; Jain, Antrix; Gordon, Aviva; Xiao, Han

doi:10.1021/acs.bioconjchem.8b00680

Cited by 64 publications

(61 citation statements)

References 35 publications

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“…[11][12][13] In parallel, site-selective chemical strategies for the conjugation of native and natural proteins have also flourishedo ver the past few years, giving rise to methods targeting varioust ypes of amino acids (for example, lysine, cysteine, tryptophan, tyrosine) that proved to be effective on proteins of all sizes, including antibodies. [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28] With the aim of pursuing the efforts in this field, we could not help but notice that the vast majority of previously reported strategies for the site-selective conjugation of native proteins were focusedo nt he modificationo faunique residue. We hypothesized that targeting two different amino acid side chainss imultaneously would lower the enormouss ubset of possibilities given by single-residue bioconjugation techniques, thus increasing our chances of developing as ite-selective method by minimising the number of potentially reactive sites;apath that has also been successfully explored by others in the meantime.…”

Section: Introductionmentioning

confidence: 99%

Investigating Ugi/Passerini Multicomponent Reactions for the Site‐Selective Conjugation of Native Trastuzumab**

Sornay

Hessmann

Erb

et al. 2020

Chemistry A European J

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Site-selective modificationo fp roteins hasb een the object of intense studies over the past decades, especially in the therapeutic field. Prominent resultsh ave been obtained with recombinant proteins,f or which site-specific conjugation is made possible by the incorporation of particular aminoa cid residues or peptides equences. In parallel, methods for the site-selective and site-specific conjugation of native andn atural proteins are starting to thrive, allowing the controlled functionalization of various types of amino acid residues.P ursuingt he efforts in this field, we planned to develop an ew typeo fs ite-selective method, aiming at the simultaneous conjugation of two amino acid residues. We reasonedt hat this should give higher chances of developing as ite-selective strategy compared to the great majority of existing methods that solely target as ingle residue. We opted for the Ugi four-centre three-component reaction to implement this idea, with the aim of conjugating the sidechain amine and carboxylate groups of two neighbouring lysine and aspartate/glutamate. Herein,w es how that this strategyc an give access to valuable antibody conjugates bearings everald ifferent payloads;f urthermore, the approach limits the potential conjugation sites to only six on the model antibody trastuzumab.

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Section: Introductionmentioning

confidence: 99%

Investigating Ugi/Passerini Multicomponent Reactions for the Site‐Selective Conjugation of Native Trastuzumab**

Sornay

Hessmann

Erb

et al. 2020

Chemistry A European J

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“…However, the lack of chemical selectivity of this method can result in nonspecific crosslinking, and 30–60 min exposure to UV irradiation has been shown to cause protein damage . To avoid the use of UV irradiation, Yu et al . and Kishimoto et al .…”

Section: Affinity Peptide Labeling For Site‐specific Conjugation Of Nmentioning

confidence: 99%

Recent Chemical Approaches for Site‐Specific Conjugation of Native Antibodies: Technologies toward Next‐Generation Antibody–Drug Conjugates

Yamada

Ito

2019

ChemBioChem

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Antibody–drug conjugates (ADCs), which consist of three components, antibody, linker, and payload, can function as “magic bullets”. These conjugates offer the ability to target drug delivery to specific cells, based on cell‐specific recognition and the binding of an antigen by a monoclonal antibody (mAb). In particular, by delivering a cytotoxic payload to cancer cells, ADCs are expected to provide a breakthrough in oncology treatments by providing a way to increase efficacy and decrease toxicity, in comparison with traditional chemotherapeutic treatments. The development of ADC therapeutics has dramatically progressed in the past decade and two ADCs have been approved and used as anticancer drugs in the clinic. However, several critical issues regarding the performance of ADCs are still being discussed and investigated. Indeed, in the past few years, several groups have reported that, changing the number and position of the drug payloads in the ADCs, affects the pharmacokinetics, drug release rates, and biological activity. The use of conventional heterogeneous conjugation methods for ADC preparation results in the drug/antibody ratio and connecting position of the payload having stochastic distributions. Therefore, it is important to investigate how these potential problems can be circumvented through site‐specific conjugation. Herein, various site‐specific chemical conjugation strategies with native mAbs that are currently used for the production of ADCs, including residue‐selective labeling for generating ADCs, disulfide rebridging, and affinity‐peptide‐mediated site‐specific chemical conjugation technologies, are reviewed and described.

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“…In addition, Furman et al [39] and Xuan et al [40] presented other reactive groups for the same purpose. All of them require the site-specific introduction of artificial amino acids [41,42], e.g., by tRNA-synthetases. This limits the applicability to genetically modified proteins [43] and may be the reason for their lack of practical use.…”

Section: Introductionmentioning

confidence: 99%

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Schroeder

Xuan

Völzke

et al. 2019

MPs

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Crosslinking of proteins for their irreversible immobilization on surfaces is a proven and popular method. However, many protocols lead to random orientation and the formation of undefined or even inactive by-products. Most concepts to obtain a more targeted conjugation or immobilization requires the recombinant modification of at least one binding partner, which is often impractical or prohibitively expensive. Here a novel method is presented, which is based on the chemical preactivation of Protein A or G with selected conventional crosslinkers. In a second step, the antibody is added, which is subsequently crosslinked in the Fc part. This leads to an oriented and covalent immobilization of the immunoglobulin with a very high yield. Protocols for Protein A and Protein G with murine and human IgG are presented. This method may be useful for the preparation of columns for affinity chromatography, immunoprecipitation, antibodies conjugated to magnetic particles, permanent and oriented immobilization of antibodies in biosensor systems, microarrays, microtitration plates or any other system, where the loss of antibodies needs to be avoided, and maximum binding capacity is desired. This method is directly applicable even to antibodies in crude cell culture supernatants, raw sera or protein-stabilized antibody preparations without any purification nor enrichment of the IgG. This new method delivered much higher signals as a traditional method and, hence, seems to be preferable in many applications.

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Proximity-Induced Site-Specific Antibody Conjugation

Cited by 64 publications

References 35 publications

Investigating Ugi/Passerini Multicomponent Reactions for the Site‐Selective Conjugation of Native Trastuzumab**

Investigating Ugi/Passerini Multicomponent Reactions for the Site‐Selective Conjugation of Native Trastuzumab**

Recent Chemical Approaches for Site‐Specific Conjugation of Native Antibodies: Technologies toward Next‐Generation Antibody–Drug Conjugates

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

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