Affimers as an alternative to antibodies for protein biomarker enrichment

Tans, Roel; Rijswijck, Danique M.H. van; Davidson, Alex; Hannam, Ryan; Ricketts, Bryon; Tack, Cees J.; Wessels, Hans; Gloerich, Jolein; Gool, Alain J. van

doi:10.1016/j.pep.2020.105677

Cited by 14 publications

(9 citation statements)

References 58 publications

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“…While antibodies are still most widely used for protein enrichment prior to (LC)-MS analysis, there are a number of examples where non-antibody-based binders were employed and sometimes also compared with antibodies. Affimers were compared to antibodies for the enrichment of the recombinant forms of two proteins, interleukin 37 (IL-37) and proinsulin, from plasma in a study by Tans et al [ 81 ]. IL-37 is a cytokine with anti-inflammatory properties, while proinsulin is the precursor of insulin, which is important for controlling blood glucose levels.…”

Section: Non-antibody-based Affinity Enrichment Of Proteins In Combination With Mass Spectrometrymentioning

confidence: 99%

Non-Antibody-Based Binders for the Enrichment of Proteins for Analysis by Mass Spectrometry

et al. 2021

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There is often a need to isolate proteins from body fluids, such as plasma or serum, prior to further analysis with (targeted) mass spectrometry. Although immunoglobulin or antibody-based binders have been successful in this regard, they possess certain disadvantages, which stimulated the development and validation of alternative, non-antibody-based binders. These binders are based on different protein scaffolds and are often selected and optimized using phage or other display technologies. This review focuses on several non-antibody-based binders in the context of enriching proteins for subsequent liquid chromatography-mass spectrometry (LC-MS) analysis and compares them to antibodies. In addition, we give a brief introduction to approaches for the immobilization of binders. The combination of non-antibody-based binders and targeted mass spectrometry is promising in areas, like regulated bioanalysis of therapeutic proteins or the quantification of biomarkers. However, the rather limited commercial availability of these binders presents a bottleneck that needs to be addressed.

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Section: Non-antibody-based Affinity Enrichment Of Proteins In Combination With Mass Spectrometrymentioning

confidence: 99%

Non-Antibody-Based Binders for the Enrichment of Proteins for Analysis by Mass Spectrometry

et al. 2021

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“…Disadvantages of using antibodies as a reagent for immunoassays include (i) long lead times for initial generation, (ii) limitations of immune responses including toxicity of potential antigens, (iii) high cost, (iv) maintenance of a consistent supply over the course of decades, and (v) stability during global shipping and long-term storage. Advances in antibody discovery and engineering such as phage display, , nanobodies, affimers, and other technologies have greatly improved some of these bottlenecks, but still leave room for improvement in particular with respect to lead times, stability, purification, and cost.…”

Section: Introductionmentioning

confidence: 99%

Development of an Aptamer-Based Electrochemical Microfluidic Device for Viral Vaccine Quantitation

et al. 2022

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Global deployment of vaccines poses significant challenges in the distribution and use of the accompanying immunoassays, one of the standard methods for quality control of vaccines, particularly when establishing assays in countries worldwide to support testing/release upon importation. This work describes our effort toward developing an integrated, portable device to carry out affinity assays for viral particles quantification in viral vaccines by incorporating (i) aptamers, (ii) microfluidic devices, and (iii) electrochemical detection. We generated and characterized more than eight aptamers against multiple membrane proteins of cytomegalovirus (CMV), which we used as a model system and designed and fabricated electrochemical microfluidic devices to measure CMV concentrations in a candidate vaccine under development. The aptamer-based assays provided a half maximal effective concentration, EC 50 , of 12 U/mL, comparable to that of an ELISA using a pair of antibodies (EC 50 60 U/mL). The device measured relative CMV concentrations accurately (within ±10% bias) and precisely (11%, percent relative standard deviation). This work represents the critical first steps toward developing simple, affordable, and robust affinity assays for global deployment without the need for sensitive equipment and extensive analyst training.

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“…The expansion of alternatives to classical antibodies led to the development of novel techniques for their manufacturing, isolation, and selection, such as phage display [ 26 ], artificial cell surface constructs and production in plants [ 27 ], generation of single-chain fragment variable (scFv) [ 28 ], among others [ 29 ]. Moreover, new types of protein binders have emerged, such as affimers, which are able to withstand a wide range of temperatures and pH levels [ 30 ], or camelid nanobodies—the smallest naturally-derived fragments, which can bind an antigen [ 31 ]. While there are more replacements for classical or recombinant antibodies and their fragments, such as aptamers [ 32 ], common immunoglobulins are still on top.…”

Section: Introductionmentioning

confidence: 99%

Recent Advancements in Receptor Layer Engineering for Applications in SPR-Based Immunodiagnostics

Drozd

Karoń

Malinowska

2021

Sensors

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The rapid progress in the development of surface plasmon resonance-based immunosensing platforms offers wide application possibilities in medical diagnostics as a label-free alternative to enzyme immunoassays. The early diagnosis of diseases or metabolic changes through the detection of biomarkers in body fluids requires methods characterized by a very good sensitivity and selectivity. In the case of the SPR technique, as well as other surface-sensitive detection strategies, the quality of the transducer-immunoreceptor interphase is crucial for maintaining the analytical reliability of an assay. In this work, an overview of general approaches to the design of functional SPR-immunoassays is presented. It covers both immunosensors, the design of which utilizes well-known and often commercially available substrates, as well as the latest solutions developed in-house. Various approaches employing chemical and passive binding, affinity-based antibody immobilization, and the introduction of nanomaterial-based surfaces are discussed. The essence of their influence on the improvement of the main analytical parameters of a given immunosensor is explained. Particular attention is paid to solutions compatible with the latest trends in the development of label-free immunosensors, such as platforms dedicated to real-time monitoring in a quasi-continuous mode, the use of in situ-generated receptor layers (elimination of the regeneration step), and biosensors using recombinant and labelled protein receptors.

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Affimers as an alternative to antibodies for protein biomarker enrichment

Cited by 14 publications

References 58 publications

Non-Antibody-Based Binders for the Enrichment of Proteins for Analysis by Mass Spectrometry

Non-Antibody-Based Binders for the Enrichment of Proteins for Analysis by Mass Spectrometry

Development of an Aptamer-Based Electrochemical Microfluidic Device for Viral Vaccine Quantitation

Recent Advancements in Receptor Layer Engineering for Applications in SPR-Based Immunodiagnostics

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