The future of protein scaffolds as affinity reagents for purification

Abstract

Affinity purification is one of the most powerful separation techniques extensively employed both at laboratory and production scales. While antibodies still represent the gold standard affinity reagents, others derived from non-immunoglobulin scaffolds emerged as interesting alternatives in particular for affinity purification. The lower costs of production, fast ligand development, and high robustness are appealing advantages of non-immunoglobulin scaffolds. These have successfully been used in the affinity purification of relevant targets as antibodies, human serum albumin, transferrin, and other biomarkers, as reviewed in this work. Furthermore, a critical assessment on the strengths, weaknesses, opportunities, and threats related with the implementation of non-immunoglobulin scaffolds as ligands in affinity purification are discussed. Biotechnol. Bioeng. 2017;114: 481-491. © 2016 Wiley Periodicals, Inc.

Affinity Sedimentation and Magnetic Separation With Plant-Made Immunosorbent Nanoparticles for Therapeutic Protein Purification

“…A range of biotic technologies beyond virus-based nanomaterials have been developed and studied for biopharmaceutical purification ( Dias and Roque, 2016 ; Mahmoodi et al, 2019 ). These can be classified by utility as fusion tags [e.g., inteins ( Wood et al, 1999 ), carbohydrate binding modules ( Shoseyov et al, 2006 )], thermo-responsive biopolymers [e.g., elastin-like polypeptides ( Sheth et al, 2014a )], and hydrophobic nanoparticles [e.g., polyhydroxyalkanoates ( Banki et al, 2005 ), oleosins ( Bhatla et al, 2010 ), hydrophobins ( Jugler et al, 2020 )].…”

Affinity sedimentation and magnetic separation with plant-made immunosorbent nanoparticles for therapeutic protein purification

“…A range of biotic technologies beyond virus-based nanomaterials have been developed and studied for biopharmaceutical purification (Dias and Roque, 2016; Mahmoodi et al , 2019). These can be classified by utility as fusion tags (e.g., inteins (Belfort et al , 1999), carbohydrate binding modules (Shoseyov et al , 2006)), thermo-responsive biopolymers (e.g., elastin-like polypeptides (Sheth, Bhut, et al , 2014)), and hydrophobic nanoparticles (e.g., polyhydroxyalkanoates (Banki et al , 2005), oleosins (Bhatla et al , 2010), hydrophobins (Jugler et al , 2020)).…”

Ways to Reach Lower Detection Limits of Lateral Flow Immunoassays

“…Thus, the single-domain antibodies produced by Tylopoda and sharks are characterized by significantly greater stability under different conditions [34]. Interest is caused by so-called protein scaffolds of a non-antibody nature that also combine conservative basic structure with hypervariable segments providing receptor functions [35]. However, their analytic application is a matter of the future.…”

“…Aptamers are significantly cheaper and more stable reagents in comparison with antibodies, and their properties are well reproducible. Limitations in affinity typical of many of the known aptamers are overcome by improving the selection procedures and subsequent directed design which is a much simpler process than for antibodies [35]. The possibilities of using aptamers in membrane test systems are shown in a number of works and summarized in recent reviews by Jauset-Rubio et al [36], Chen et al [37], and Dhiman et al [38].…”