Disulfide Scrambling in IgG2 Monoclonal Antibodies: Insights from Molecular Dynamics Simulations

Wang, Xiaoling; Kumar, Sandeep; Singh, Satish K.

doi:10.1007/s11095-011-0503-9

Cited by 40 publications

(39 citation statements)

References 33 publications

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“…17 Molecular dynamic simulation study revealed that the sulfur atoms of inter-chain disulfide bonds are highly mobile and can be in close proximity. 18 Therefore, it is not a surprise to observe the coexistence of multiple disulfide bond isoforms for IgG 2 antibodies. In addition to isoforms from different intramolecule disulfide bond linkages, disulfide bond linked IgG 2 dimer was also found in recombinant IgG 2 from cell culture and in human serum.…”

Section: Free Sulfhydrylsmentioning

confidence: 99%

Disulfide bond structures of IgG molecules

Liu

May

2012

mAbs

372

204

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Section: Free Sulfhydrylsmentioning

confidence: 99%

Disulfide bond structures of IgG molecules

Liu

May

2012

mAbs

372

204

View full text Add to dashboard Cite

“…Although the isoforms only differ in the arrangement of the inter-chain disulfide bonds, each disulfide pattern alters the higher-order structure of the mAb, thereby affecting thermal stability, binding efficiency, surface hydrophobicity, isoelectric point, and possibly influencing developability. 15–19 For example, A-isoform enriched panitumumab and denosumab demonstrated higher apparent affinity to the human IgG Fc receptor-like 5 (FCRL5) compared to the B isoform. 20 Disulfide isoform distribution may also affect the conjugation profile of cysteine-linked antibody-drug conjugates.…”

Section: Introductionmentioning

confidence: 99%

“…17,20,22–26 The flexible and solvent-accessible hinge region is particularly crucial, as it tends to be labile to reduction and oxidation that can lead to disulfide scrambling or trisulfide formation. 15,17,19,27–30 Several mass spectrometry (MS)/MS approaches for characterizing disulfide linkages have been described using collision-induced dissociation (CID), electron-transfer dissociation (ETD), or electron-capture dissociation (ECD). 11–13,29,31–35 These approaches often require multiple analysis, comparison of reduced and non-reduced peptide maps, interpretation of complex data sets, or large sample consumption.…”

Section: Introductionmentioning

confidence: 99%

Rapid, automated characterization of disulfide bond scrambling and IgG2 isoform determination

Resemann

Liu-Shin

Tremintin

et al. 2018

mAbs

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Human antibodies of the IgG2 subclass exhibit complex inter-chain disulfide bonding patterns that result in three structures, namely A, A/B, and B. In therapeutic applications, the distribution of disulfide isoforms is a critical product quality attribute because each configuration affects higher order structure, stability, isoelectric point, and antigen binding. The current standard for quantification of IgG2 disulfide isoform distribution is based on chromatographic or electrophoretic techniques that require additional characterization using mass spectrometry (MS)-based methods to confirm disulfide linkages. Detailed characterization of the IgG2 disulfide linkages often involve MS/MS approaches that include electrospray ionization or electron-transfer dissociation, and method optimization is often cumbersome due to the large size and heterogeneity of the disulfide-bonded peptides. As reported here, we developed a rapid LC-MALDI-TOF/TOF workflow that can both identify the IgG2 disulfide linkages and provide a semi-quantitative assessment of the distribution of the disulfide isoforms. We established signature disulfide-bonded IgG2 hinge peptides that correspond to the A, A/B, and B disulfide isoforms and can be applied to the fast classification of IgG2 isoforms in heterogeneous mixtures.

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“…Hence, computer simulation has become an indispensable tool to study the shape, size, conformational stability, hydrodynamic behavior, folding patterns, and denaturation of such peptides. [24][25][26] So far, about 500 different types of cone snail species are reported. The pioneering work of characterization of cone snail venoms was done by Spence et al 27 They studied the venom from Conus geographus, purified it and attempted to determine its amino acid composition.…”

Section: Introductionmentioning

confidence: 99%

“…Though coarse in nature, this methodology has been used before by computation chemists to characterize the higher order structures in proteins and other biomolecules with reasonable accuracy. 12,26 In D, both the native disulfide bonds are broken and so the reasonably labile peptide is free to explore the energy landscape by acquiring different conformations. Figure 7 shows the distribution of the distances for the three possible combinations of the four cysteine residues to form the two disulfide bonds.…”

mentioning

confidence: 99%

Scrambling of disulfide bond scaffolds in neurotoxin AuIB: A molecular dynamics simulation study

Roy

Lakshminarayanan

2016

Biopolymers

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AuIB, a neurotoxic conopeptide, is a "selective antagonist" of the α3-β4 subtype of the nicotinic acetylcholine receptors in human brain and is investigated extensively for its immense potency in the treatment of pain. It contains two disulfide linkages, which are decisive in maintaining its structure and functional selectivity. Here we report, a molecular dynamics simulation study of the role of disulfide bonds on the secondary structure of AuIB in water. The native form of AuIB (N) is found to be significantly stable in water with very robust 3 and α-helical domains, featuring ∼47% of the total structure. The partially reduced AuIB (P ), with the disulfide bond between cysteine 2, 8 broken, shows significantly perturbed secondary structural features with almost total loss in helicity. Breaking of the disulfide bond between cysteine 3, 15 (P ), on the other hand, has almost no effect on the helical region of the peptide, although the weightage of β-turn increased at the cost of random coil. Intriguingly, when both the disulfide bonds are broken (D), the helical region is affected, but the loss in helicity is less than that observed in the P case. To understand the disulfide scrambling process, the relative probabilities of forming three disulfide-bond isoforms of AuIB in water are estimated from the sulfur-sulfur (SS) distance distributions of the four cysteine residues present in AuIB (D). Simulations show that the native globular form dominates ∼73% of the isoform population, with the beads form being the second most populated one. © 2015 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 196-209, 2016.

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Disulfide Scrambling in IgG2 Monoclonal Antibodies: Insights from Molecular Dynamics Simulations

Abstract: Consistent with experimental studies, our simulations indicate inter-chain disulfide linkages in human IgG2 mAbs are degenerate. Potential rational design strategies to devise hinge stabilized human IgG2 mAbs are gleaned.

Cited by 40 publications

References 33 publications

Disulfide bond structures of IgG molecules

Disulfide bond structures of IgG molecules

Rapid, automated characterization of disulfide bond scrambling and IgG2 isoform determination

Scrambling of disulfide bond scaffolds in neurotoxin AuIB: A molecular dynamics simulation study

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