Light-induced aggregation of type I soluble tumor necrosis factor receptor

Roy, Shouvik; Mason, Bruce D.; Schöneich, Christian; Carpenter, John F.; Boone, Thomas C.; Kerwin, Bruce A.

doi:10.1002/jps.21750

Cited by 27 publications

(34 citation statements)

References 68 publications

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“…Thermal unfolding by monitoring changes in tryptophan fluorescence was also used to study the stability of the In direct contrast to what we observed for anti-streptavidin, the tertiary structural analysis of sTNF-R1 exposed to 302 nm light showed changes in the positioning of the tryptophan and phenylalanine residues in asymmetric environments. 39 For anti-streptavidin exposed to light, a close examination of the tertiary structure by near UV CD as well as 2 nd derivative UV-vis spectroscopy and intrinsic tryptophan fluorescence at 25ºC failed to detect significant differences at pH 5 and 8 upon exposure under the conditions described above. At pH 3.5, there may be a small difference for the 1 mg/mL sample as observed by near UV CD.…”

Section: Effect Of Light On Conformation Of the Anti-streptavidin Mabmentioning

confidence: 96%

“…30 For comparison, the light-induced cleavage of glutathione disulfide led predominantly to thiyl radicals. 30 It is well documented that radical mediated damage to proteins may lead to a variety of potentially negative consequences such as fragmentation 23,38 , aggregation 25,38,39 , oxidation [40][41][42][43] , changes to surface hydrophobicity, conformational changes 39 and even denaturation. 38,39 It may also ultimately result in the loss of activity or simply generate new reactive species propagating the radical reactions.…”

Section: Chemistry Of Photooxidationmentioning

confidence: 99%

“…followed by emission scan at 440-450nm) 39 was negative as was a fluorescence test for dityrosine 60 (data not shown). This is in contrast to sTNF-R1, which formed Nformylkynurenine upon exposure to 302nm light, implicating its sole tryptophan in the degradation process.…”

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confidence: 95%

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Effect of pH and Light on Aggregation and Conformation of an IgG1 mAb

2012

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Abstract:During the purification process, monoclonal antibodies may be exposed to parts of UV-C (200 to 290 nm), UV-B (290 to 320 nm) and visible light (400 to 760 nm) under a variety of buffer and pH conditions. Together, these conditions can promote both chemical and physical degradation which may result in conformational changes. To examine this possibility, the impact of UV light on an IgG1 mAb at pH 3.5, 5 and 8 was studied at multiple protein concentrations. Exposure to 302 nm light resulted in a pH-dependent formation of high molecular weight species where the degree of oligomerization increased with increasing pH.Characterization by SDS-PAGE under reducing and non-reducing conditions, and SEC-MALS, revealed that the predominant species were non-reducible dimeric, trimeric and higher order oligomeric species which occurred through processes other than intermolecular disulfide bond showing the greatest degree of change in the β-sheet structure. Exposure to UV light resulted in aggregation with pH-dependent yields decreasing in the following order, 8.0>5.0>3.5, while the opposite trend was observed for conformational changes, with pH-dependent extents decreasing in the following order:3.5>5.0>8.0. These pH-dependent trends suggest that different strategies will be required to stabilize the protein against these modifications during processing.

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Section: Effect Of Light On Conformation Of the Anti-streptavidin Mabmentioning

confidence: 96%

Section: Chemistry Of Photooxidationmentioning

confidence: 99%

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Effect of pH and Light on Aggregation and Conformation of an IgG1 mAb

2012

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“…When Trp is photoexcited by absorbing near-UV light, it can affect neighboring amino acids and, in doing so, can reduce disulfide bonds (193,223). The photoionization of Trp can reduce disulfide bonds by electron transfer, resulting in chemical and physical degradation of the protein (193,(223)(224)(225).…”

Section: Photooxidationmentioning

confidence: 98%

“…Intrinsic actors include the flexibility of the peptide backbone (188) and the overall structure of the protein (188)(189)(190). In addition, extrinsic factors, such as pH (191)(192)(193) and buffer type (191,194), can affect oxidation rates of proteins as well.…”

Section: Oxidationmentioning

confidence: 99%

Stability of Protein Pharmaceuticals: An Update

Manning¹,

Chou²,

Murphy³

et al. 2010

Pharm Res

999

782

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In 1989, Manning, Patel, and Borchardt wrote a review of protein stability (Manning et al., Pharm. Res. 6:903-918, 1989), which has been widely referenced ever since. At the time, recombinant protein therapy was still in its infancy. This review summarizes the advances that have been made since then regarding protein stabilization and formulation. In addition to a discussion of the current understanding of chemical and physical instability, sections are included on stabilization in aqueous solution and the dried state, the use of chemical modification and mutagenesis to improve stability, and the interrelationship between chemical and physical instability.

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