Reduction of biological activity of murine recombinant interleukin‐1β by selective deamidation at asparagine‐149

Daumy, Gaston O.; Wilder, Cheryl; Merenda, Joseph M.; McColl, A S; Geoghegan, Kieran F.; Otterness, Ivan G.

doi:10.1016/0014-5793(91)80093-i

Cited by 19 publications

(1 citation statement)

References 30 publications

(1 reference statement)

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“…Preparation of Protein Reagents . Recombinant wild-type human IL-1β (hIL-1β), hIL-1β muteins, and recombinant wild-type murine IL-1α and -1β (mIL-1α and mIL-1β) were prepared as described ( − ). The soluble recombinant human IL-1 receptor (sIL-1R) and wild-type human IL-1α (hIL-1α) protein preparations were purchased from Genzyme (Cambridge, MA) and Collaborative, Research (Bedford, MA), respectively.…”

Section: Methodsmentioning

confidence: 99%

Inhibiting Protein−Protein Interactions: A Model for Antagonist Design

Chrunyk

Rosner²,

Yao³

et al. 2000

Biochemistry

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Protein-protein interactions (PPI) are a ubiquitous mode of transmitting signals in cells and tissues. We are testing a stepwise, generic, structure-driven approach for finding low molecular weight inhibitors of protein-protein interactions. The approach requires development of a high-affinity, single chain antibody directed specifically against the interaction surface of one of the proteins to obtain structural information on the interface. To this end, we developed a single chain antibody (sc1E3) against hIL-1beta that exhibited the equivalent affinity of the soluble IL-1 receptor type I (sIL-1R) for hIL-1beta and competitively blocked the sIL-1R from binding to the cytokine. The antibody proved to be more specific for hIL-1beta than the sIL-1R in that it failed to bind to either murine IL-1beta or human/murine IL-1alpha proteins. Additionally, failure of sc1E3 to bind to several hIL-1beta mutant proteins, altered at receptor site B, indicated that the antibody interacted preferentially with this site. This, coupled with other surface plasmon resonance and isothermal titration calorimetry measurements, shows that sc1E3 can achieve comparable affinity of binding hIL-1beta as the receptor through interactions at a smaller interface. This stable single chain antibody based heterodimer has simplified the complexity of the IL-1/IL-1R PPI system and will facilitate the design of the low molecular weight inhibitors of this interaction.

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Section: Methodsmentioning

confidence: 99%

Inhibiting Protein−Protein Interactions: A Model for Antagonist Design

Chrunyk

Rosner²,

Yao³

et al. 2000

Biochemistry

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Quality attributes of recombinant therapeutic proteins: An assessment of impact on safety and efficacy as part of a quality by design development approach

Eon‐Duval¹,

Broly²,

Gleixner³

2012

Biotechnology Progress

176

123

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Quality by Design (QbD) is a new approach to the development of recombinant therapeutic protein products that promotes a better understanding of the product and its manufacturing process. The first step in the QbD approach consists in identifying the critical quality attributes (CQA), i.e., those quality attributes of the product that have an impact on its clinical efficacy or safety. CQAs are identified through a science-based risk assessment taking into consideration a combination of clinical and nonclinical data obtained with the molecule or other similar molecules or platform products, as well as the published literature. The purpose of this article is to perform a comprehensive review of the published literature, supporting an assessment of the impact on safety and efficacy of the quality attributes commonly encountered in recombinant therapeutic proteins, more specifically those produced in mammalian cell expression systems. Quality attributes generally observed in biopharmaceutical proteins including product-related impurities and substances, process-related impurities, product attributes, and contaminants are evaluated one by one for their impact on biological activity, pharmacokinetics and pharmacodynamics, immunogenicity, and overall safety/toxicity.

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Purification and preliminary crystallographic studies of penicillin G acylase from Providencia rettgeri

1995

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Two isoforms of the heterodimeric enzyme penicillin G acylase (EC 3.5.1.1 I ) from Providencia rettgeri ATCC 31052 (strain Brol) were purified to near homogeneity. The isoforms exhibited comparable enzymatic activities but differed slightly in the molecular weight and PI of their respective a-subunit. The origin of this difference was traced to the partial conversion of the N-terminal Gln of the a-subunit to pyrrolidonecarboxylic acid (pyro-Glu). The boundaries of the mature enzyme within the translated DNA sequence of the wild-type propeptide (GenBank M86533) were determined. The results conclusively identified the length of the signal peptide and the position of the spacer cleaved from the propeptide to form the active heterodimer. The molecular weights of the cy-and 0-subunits, based on these termini, were 23.7 and 62.2 kDa, respectively. Both isoforms were crystallized independently as hexagonal bipyramids up to 0.60 mm in diameter in either space group P6,22 or P6522 ( a = b = 140.5 A and c = 209.5 A) from ammonium sulfate solutions buffered by 50 mM potassium phosphate at pH 7.5.The presence of glycerol, although not required, facilitated crystal growth. Native and heavy atom derivative data were collected to 3.0 A resolution, and the calculation of isomorphous replacement phases is under way.

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Reduction of biological activity of murine recombinant interleukin‐1β by selective deamidation at asparagine‐149

Cited by 19 publications

References 30 publications

Inhibiting Protein−Protein Interactions: A Model for Antagonist Design

Inhibiting Protein−Protein Interactions: A Model for Antagonist Design

Quality attributes of recombinant therapeutic proteins: An assessment of impact on safety and efficacy as part of a quality by design development approach

Purification and preliminary crystallographic studies of penicillin G acylase from Providencia rettgeri

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