Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G.

Eliasson, Margareta; Olsson, Anders; Palmcrantz, Elisabeth; Wiberg, Kristina; Inganäs, M.; Guss, Bengt; Lindberg, Martin; Uhlén, Mathias

doi:10.1016/s0021-9258(18)68928-8

Cited by 166 publications

(48 citation statements)

References 19 publications

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“…Protein G reportedly exhibits broader species specificity with less variation in isotype binding within a species than other nonspecies-specific Ig-binding cell wall proteins such as protein A. 5 Many investigators also report stronger binding affinity between protein G and Ig from several species, 2,3,5,9,10,14 as well as selective reactivity to IgG. 5,12,14,24 Reports on the binding of Fc-receptor bacterial proteins to sera of domestic and wild African mammals, 16 birds, and reptiles 18 found that whole sera from large ungulates (e.g., wildebeest, waterbuck, buffalo, nyala) reacted strongly with the chimeric protein (containing recombinant protein G and protein A) and weakly with protein A alone.…”

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Protein G Binding to Enriched Serum Immunoglobulin from Nondomestic Hoofstock Species

2003

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Abstract. Quick and cost-effective serologic assays, such as those based on enzyme-linked immunosorbent assay (ELISA) technology, are useful for screening animal populations for infectious diseases. Recombinant protein G is described as an almost universal ELISA conjugate for the detection of antibodies from a wide range of animal species. However, there is limited data documenting the ability of protein G to bind immunoglobulin (Ig) from many captive and free-ranging nondomestic hoofstock (Order Artiodactyla, e.g., elk, antelope, bison). Protein G binding to Ig from 11 species within this taxonomic order (addax, antelope, bison, bontebok, elk, impala, kudu/nyala, muntjac, oryx, sheep, and white-tailed deer) and 2 control species (bovine and chicken) was assessed. A serum Ig enrichment protocol, using high-performance liquid chromatography (HPLC), was optimized in bovids (Bos taurus) and then applied to the other study species. Binding assays were performed by adding protein G to microtiter wells coated with titrated dilutions of enriched artiodactyl Ig. Optical densities were measured and binding curves generated. Differences in protein G binding were observed, both within and among species, as well as within taxonomic families. Significant intraspecies binding variation was observed for 7 species tested (antelope, oryx, sheep, muntjac, impala, bontebok, and addax). No statistically significant intraspecies differences in protein G binding were found for Ig from bison, elk, kudu/nyala, whitetailed deer, plus control species (cattle and chicken). Binding of protein G to Ig from impala, muntjac, and elk was statistically different from the positive control (cattle), with muntjac binding curves statistically comparable with the negative control (chicken). For the other 7 species tested, binding curves illustrated the ability of protein G to bind Ig as well as, or better than, the positive control. These findings expand the list of animal species whose Ig is capable of being detected using recombinant protein G, with the caveat that protein G does not bind Ig uniformly in closely related species. It is concluded that recombinant protein G conjugates may serve as useful reagents for serodiagnosis by ELISA in nondomestic hoofstock, although different assay interpretation algorithms and assay protocols may need to be developed on a per species basis for maximum diagnostic effectiveness.Detection, control, and management of infectious diseases in captive and free-ranging nondomestic ruminant hoofstock species (Order Artiodactyla) often prove challenging. These species, such as okapi, impala, and bontebok, are not often handled, and as a result, samples for diagnostic analysis are infrequently collected. An additional hindrance to disease surveillance is the lack of validated diagnostic assays for these valuable, and in some cases, endangered species.Reliable, cost-effective, and noninvasive diagnostics are needed to maximize the value of diagnostic samples when they can be collected. In particular, for diseases eliciti...

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Protein G Binding to Enriched Serum Immunoglobulin from Nondomestic Hoofstock Species

2003

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“…Proteins A, G, and L have a scope of biochemical, biotechnological, and medical applications and are widely used in antibody purification and test methods like immunoprecipitation, ELISA, and Western blotting [ 153 ]. The “universal” recombinant fusion protein A/G comprised of four domains of protein A and two domains of protein G demonstrates additive properties of the two proteins and captures antibodies from the combined spectra of species and isotypes [ 154 , 155 ].…”

Section: Engineered Protein G and Antibody-scaffold Variantsmentioning

confidence: 99%

Protein Engineering: Advances in Phage Display for Basic Science and Medical Research

Davydova

2022

Biochemistry Moscow

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Functional Protein Engineering became the hallmark in biomolecule manipulation in the new millennium, building on and surpassing the underlying structural DNA manipulation and recombination techniques developed and employed in the last decades of 20th century. Because of their prominence in almost all biological processes, proteins represent extremely important targets for engineering enhanced or altered properties that can lead to improvements exploitable in healthcare, medicine, research, biotechnology, and industry. Synthetic protein structures and functions can now be designed on a computer and/or evolved using molecular display or directed evolution methods in the laboratory. This review will focus on the recent trends in protein engineering and the impact of this technology on recent progress in science, cancer- and immunotherapies, with the emphasis on the current achievements in basic protein research using synthetic antibody (sABs) produced by phage display pipeline in the Kossiakoff laboratory at the University of Chicago (KossLab). Finally, engineering of the highly specific binding modules, such as variants of Streptococcal protein G with ultra-high orthogonal affinity for natural and engineered antibody scaffolds, and their possible applications as a plug-and-play platform for research and immunotherapy will be described.

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“…Comparison with protein A and G coupled to HRP from two different vendors using serum from numerous species demonstrated that SA-TRIM21 is a multispecies detection probe with broader specificity than traditional Protein A and G detection probes (Figure S3). Since bacterial proteins A and G have complementary binding patterns, a fusion protein enclosing both proteins generated broader reporter molecules (Eliasson et al, 1988;Sun & Lew, 1992). More recently, engineered proteins were selected to recognize different epitopes within human IgG and exhibited binding affinities in the nanomolar range (Behar et al, 2013).…”

Section: Igg and Animal Serum Detection Using Sa-trim21mentioning

confidence: 99%

TRIM21 chimeric protein as a new molecular tool for multispecies IgG detection

Ramos

Fernandes

Batista

et al. 2021

Preprint

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G-type immunoglobulins (IgGs) are extensively used in the pharmaceutical industry against various diseases, being also crucial in multiple immunoassays. The production of secondary monoclonal antibodies (Abs) for IgG detection is not cost-effective, while polyclonal antibody production still depends on laboratory animals, which raises concerns regarding animal welfare. As alternatives, bacterial proteins (A and G) have been widely exploited; however, several difficulties are encountered regarding their use for IgG detection and purification. The widespread use of IgGs in the pharmaceutical industry and the increasing number and variety of new Abs entering the market impose the need to develop new detection and purification strategies. The TRIM21 protein is a soluble intracellular IgG receptor that binds to the Fc region of many species with high affinity. We created a chimeric protein containing a mutated form of the C-terminal domain of mouse TRIM21 linked to a streptavidin moiety to detect IgGs from a wide range of species. The protein is promptly produced by heterologous expression and consists of an improved molecular tool, expanding the portfolio of Ab-affinity ligands for immunoassays.

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Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G.

Cited by 166 publications

References 19 publications

Protein G Binding to Enriched Serum Immunoglobulin from Nondomestic Hoofstock Species

Protein G Binding to Enriched Serum Immunoglobulin from Nondomestic Hoofstock Species

Protein Engineering: Advances in Phage Display for Basic Science and Medical Research

TRIM21 chimeric protein as a new molecular tool for multispecies IgG detection

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