CovX‐Bodies

Bhat, Abhijit; and, Olivier Laurent; Lappe, Rodney W.

doi:10.1002/9781118354599.ch38

Cited by 5 publications

(12 citation statements)

References 30 publications

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“…In contrast to any conjugation technology previously described, the antibody acts here solely as a carrier protein and has no targeting properties. By fusing a small molecular weight payload (SMWP, e.g., peptide) with a catalytic antibody, it is possible to generate antibody conjugates that combine the highly specific targeting properties of the SMWP and the pharmacokinetic characteristics of an antibody [168][169][170]. The conjugation is based on the derivatization of the pharmacophore with either a diketone, vinylketone or azetidinone functional group that forms a covalent bond with a specific lysine residue in the active site of the catalytic antibody ( Figure 8B).…”

Section: Catalytic Antibodiesmentioning

confidence: 99%

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

2015

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Monoclonal antibodies (mAbs) and their derivatives are currently the fastest growing class of therapeutics. Even if naked antibodies have proven their value as successful biopharmaceuticals, they suffer from some limitations. To overcome suboptimal therapeutic efficacy, immunoglobulins are conjugated with toxic payloads to form antibody drug conjugates (ADCs) and with chelating systems bearing therapeutic radioisotopes to form radioimmunoconjugates (RICs). Besides their therapeutic applications, antibody conjugates are also extensively used for many in vitro assays. A broad variety of methods to functionalize antibodies with various payloads are currently available. The decision as to which conjugation method to use strongly depends on the final purpose of the antibody conjugate. Classical conjugation via amino acid residues is still the most common method to produce antibody conjugates and is suitable for most in vitro applications. In recent years, however, it has become evident that antibody conjugates, which are generated via site-specific conjugation techniques, possess distinct advantages with regard to in vivo properties. Here, we give a comprehensive overview on existing and emerging strategies for the production of covalent and non-covalent antibody conjugates.

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Section: Catalytic Antibodiesmentioning

confidence: 99%

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

2015

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“…The inherent complexity of these bioconjugates can pose significant hurdles to late‐stage process development. To illustrate this, we report for the first time key late‐stage process improvements for a representative complex bioconjugate, where the CovX half‐life extension technology that uses an antibody scaffold for peptide and small protein therapeutics, is applied to Fibroblast Growth Factor 21 (FGF21).…”

Section: Introductionmentioning

confidence: 99%

“…The FGF21‐antibody conjugate, CVX‐343, is an extended half‐life therapeutic that has reached Phase 1 Clinical Trials . The conjugate consists of three components: a recombinant FGF21 cysteine (Cys) mutant protein (FGF21 A129C ) produced in E. coli , an aldolase monoclonal antibody (mAb) scaffold for half‐life extension produced in Chinese Hamster Ovary (CHO) cells, and a chemically synthesized heterobifunctional, maleimide‐PEG 2 ‐azetidinone linker (mal‐PEG 2 ‐AZD). The linker is equipped with a maleimide (mal) group for reaction with FGF21 A129C and an azetidinone (AZD) group for conjugation to the mAb scaffold.…”

Section: Introductionmentioning

confidence: 99%

“…The linker is equipped with a maleimide (mal) group for reaction with FGF21 A129C and an azetidinone (AZD) group for conjugation to the mAb scaffold. The aldolase mAb scaffold has a catalytic pocket located on each of its heavy chains with a uniquely reactive lysine (Lys) that can be targeted for site‐selective conjugation . The linker is designed such that it binds into these pockets positioning the AZD group for reaction with the Lys side chain amine to give a stable amide linkage .…”

Section: Introductionmentioning

confidence: 99%

“…The aldolase mAb scaffold has a catalytic pocket located on each of its heavy chains with a uniquely reactive lysine (Lys) that can be targeted for site‐selective conjugation . The linker is designed such that it binds into these pockets positioning the AZD group for reaction with the Lys side chain amine to give a stable amide linkage . As a result, a defined number of two FGF21 A129C proteins can be site‐selectively conjugated to the mAb scaffold.…”

Section: Introductionmentioning

confidence: 99%

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Process development of a FGF21 protein–antibody conjugate

et al. 2017

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A scalable, viable process was developed for the Fibroblast Growth Factor 21 (FGF21) proteinantibody conjugate, CVX-343, an extended half-life therapeutic for the treatment of metabolic disease. CVX-343 utilizes the CovX antibody scaffold technology platform that was specifically developed for peptide and protein half-life extension. CVX-343 is representative of a growing number of complex novel peptide-and protein-based bioconjugate molecules currently being explored as therapeutic candidates. The complexity of these bioconjugates, assembled using wellestablished chemistries, can lead to very difficult production schemes requiring multiple starting materials and a combination of diverse technologies. Key improvements had to be made to the original CVX-343 Phase 1 manufacturing process in preparation for Phase 3 and commercial manufacturing. A strategy of minimizing FGF21 A129C dimerization and stabilizing the FGF21 A129C Drug Substance Intermediate (DSI), linker, and activated FGF21 intermediate was pursued. The use of tris(2-carboxyethyl)phosphine (TCEP) to prevent FGF21 A129C dimerization through disulfide formation was eliminated. FGF21 A129C dimerization and linker hydrolysis were minimized by formulating and activating FGF21 A129C at acidic instead of neutral pH. An activation use test was utilized to guide FGF21 A129C pooling in order to minimize misfolds, dimers, and misfolded dimers in the FGF21 A129C DSI. After final optimization of reaction conditions, a process was established that reduced the consumption of FGF21 A129C by 36% (from 4.7 to 3.0 equivalents) and the consumption of linker by 55% (from 1.4 to 0.95 equivalents for a smaller required amount of FGF21 A129C ). The overall process time was reduced from 5 to 3 days. The product distribution improved from containing 60% to 75% desired bifunctionalized (12 FGF21) FGF21-antibody conjugate in the crude conjugation mixture and from 80% to 85% in the final CVX-343 Drug Substance (DS), while maintaining the same overall process yield based on antibody scaffold input. FGF21, a 19 kDa secreted protein, is a modulator of glucose, lipid, and energy homeostasis and has gained significant interest as a therapeutic for treating metabolic disease. [9][10][11][12][13][14] However, its short plasma half-life, [9,15,16] limited proteolytic stability, [9,17,18] and tendency to aggregate [9,17,18] have posed significant challenges to the development of a FGF21 therapeutic. Various protein engineering and conjugation approaches have been taken to address these limitations. [9] Among these are strategies to extend the half-life of FGF21 through PEGylation, [19] Fc fusion, [20] or conjugation to an antibody scaffold. [7] The FGF21-antibody conjugate, CVX-343, is an extended half-life therapeutic that has reached Phase 1 Clinical Trials. [7,[21][22][23] The conjugate consists of three components: a recombinant FGF21 cysteine (Cys) mutant protein (FGF21 A129C ) produced in E. coli, an aldolase monoclonal antibody (mAb) scaffold for half-life extension produced...

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Fusion Proteins for Half-Life Extension of Biologics as a Strategy to Make Biobetters

2015

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The purpose of making a “biobetter” biologic is to improve on the salient characteristics of a known biologic for which there is, minimally, clinical proof of concept or, maximally, marketed product data. There already are several examples in which second-generation or biobetter biologics have been generated by improving the pharmacokinetic properties of an innovative drug, including Neulasta® [a PEGylated, longer-half-life version of Neupogen® (filgrastim)] and Aranesp® [a longer-half-life version of Epogen® (epoetin-α)]. This review describes the use of protein fusion technologies such as Fc fusion proteins, fusion to human serum albumin, fusion to carboxy-terminal peptide, and other polypeptide fusion approaches to make biobetter drugs with more desirable pharmacokinetic profiles.

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CovX‐Bodies

Cited by 5 publications

References 30 publications

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

Antibody Conjugates: From Heterogeneous Populations to Defined Reagents

Process development of a FGF21 protein–antibody conjugate

Fusion Proteins for Half-Life Extension of Biologics as a Strategy to Make Biobetters

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