A capture and release method based on noncovalent ligand cross-linking and facile filtration for purification of lectins and glycoproteins

Welch, Christina; Talaga, Melanie L.; Kadav, Priyanka; Edwards, Jared L.; Bandyopadhyay, Purnima; Dam, Tarun K.

doi:10.1074/jbc.ra119.010625

Cited by 12 publications

(14 citation statements)

References 45 publications

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“…Signal-stimulated release of (bio)molecules is a challenging goal [1][2][3] and the high research activity in this area is justified by many important practical applications. 4,5 The major interest in these studies is directed to the controlled release of drugs in medical applications, 6,7 however, some other applications, for example, in separation and purification of substances 8,9 are also feasible. Many different systems have been researched for the controlled release based on a broad variety of the stimulating input signals (light irradiation, 10 change of temperature 11 or pH values, 12 applications of magnetic field 13 or electric potential, 14 additions of (bio)chemical substances, 15 etc.).…”

mentioning

confidence: 99%

Electrochemically produced local pH changes stimulating (bio)molecule release from pH-switchable electrode-immobilized avidin–biotin systems

Badenhorst

Kadambar

Bellare

et al. 2022

Phys. Chem. Chem. Phys.

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mentioning

confidence: 99%

Electrochemically produced local pH changes stimulating (bio)molecule release from pH-switchable electrode-immobilized avidin–biotin systems

Badenhorst

Kadambar

Bellare

et al. 2022

Phys. Chem. Chem. Phys.

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“…One technology in particular is the combination of solid-phase extraction methods, using affinity-based ligands, with CE. Highly selective affinity ligands or adsorbents, such as antibodies, antibody fragments, lectins, aptamers, enzymes, metal-organic framework-based affinity sorbents, and other specially designed ligands are the cornerstone for the isolation and purification of many substances and cellular-subcellular entities found in complex matrices [ 25 , 47 , 54 , 55 , 56 , 57 , 58 , 75 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 ].…”

Section: Role Of Capillary Electrophoresis In Point-of-care Diagnomentioning

confidence: 99%

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

Guzman¹,

Guzman

2020

Biomedicines

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Biomarker detection for disease diagnosis, prognosis, and therapeutic response is becoming increasingly reliable and accessible. Particularly, the identification of circulating cell-free chemical and biochemical substances, cellular and subcellular entities, and extracellular vesicles has demonstrated promising applications in understanding the physiologic and pathologic conditions of an individual. Traditionally, tissue biopsy has been the gold standard for the diagnosis of many diseases, especially cancer. More recently, liquid biopsy for biomarker detection has emerged as a non-invasive or minimally invasive and less costly method for diagnosis of both cancerous and non-cancerous diseases, while also offering information on the progression or improvement of disease. Unfortunately, the standardization of analytical methods to isolate and quantify circulating cells and extracellular vesicles, as well as their extracted biochemical constituents, is still cumbersome, time-consuming, and expensive. To address these limitations, we have developed a prototype of a portable, miniaturized instrument that uses immunoaffinity capillary electrophoresis (IACE) to isolate, concentrate, and analyze cell-free biomarkers and/or tissue or cell extracts present in biological fluids. Isolation and concentration of analytes is accomplished through binding to one or more biorecognition affinity ligands immobilized to a solid support, while separation and analysis are achieved by high-resolution capillary electrophoresis (CE) coupled to one or more detectors. When compared to other existing methods, the process of this affinity capture, enrichment, release, and separation of one or a panel of biomarkers can be carried out on-line with the advantages of being rapid, automated, and cost-effective. Additionally, it has the potential to demonstrate high analytical sensitivity, specificity, and selectivity. As the potential of liquid biopsy grows, so too does the demand for technical advances. In this review, we therefore discuss applications and limitations of liquid biopsy and hope to introduce the idea that our affinity capture-separation device could be used as a form of point-of-care (POC) diagnostic technology to isolate, concentrate, and analyze circulating cells, extracellular vesicles, and viruses.

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“…Therefore, binding affinity of the targets for selected multivalent ligands (potential TCAs) is determined by hemagglutination inhibition assays. The most commonly tested multivalent ligands included invertase, bovine thyroglobulin, mannan, asialofetuin, and chondroitin sulfate A and C (see Welch et al, 2020). A representative inhibition profile is shown in Figure 5.…”

Section: Identification Of Multivalent Inhibitors As Target Capturingmentioning

confidence: 99%

“…The present capture and release (CaRe) method was formulated to address some of these issues. CaRe was successfully employed to purify several lectins and glycoproteins (see Welch et al, 2020). CaRe is faster and cheaper.…”

Section: Of 27mentioning

confidence: 99%

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A Rapid and Facile Purification Method for Glycan‐Binding Proteins and Glycoproteins

et al. 2020

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Glycosylated proteins, namely glycoproteins and proteoglycans (collectively called glycoconjugates), are indispensable in a variety of biological processes. The functions of many glycoconjugates are regulated by their interactions with another group of proteins known as lectins. In order to understand the biological functions of lectins and their glycosylated binding partners, one must obtain these proteins in pure form. The conventional protein purification methods often require long times, elaborate infrastructure, costly reagents, and large sample volumes. To minimize some of these problems, we recently developed and validated a new method termed capture and release (CaRe). This method is time‐saving, precise, inexpensive, and it needs a relatively small sample volume. In this approach, targets (lectins and glycoproteins) are captured in solution by multivalent ligands called target capturing agents (TCAs). The captured targets are then released and separated from their TCAs to obtain purified targets. Application of the CaRe method could play an important role in discovering new lectins and glycoconjugates. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Preparation of crude extracts containing the target proteins from soybean flour Alternate Protocol 1: Preparation of crude extracts from Jack bean meal Alternate Protocol 2: Preparation of crude extracts from the corms of Colocasia esculenta, Xanthosoma sagittifolium, and from the bulbs of Allium sativum Alternate Protocol 3: Preparation of Escherichia coli cell lysates containing human galectin‐3 Alternate Protocol 4: Preparation of crude extracts from chicken egg whites (source of ovalbumin) Basic Protocol 2: Preparation of 2% (v/v) red blood cell suspension Basic Protocol 3: Detection of lectin activity of the crude extracts Basic Protocol 4: Identification of multivalent inhibitors as target capturing agents by hemagglutination inhibition assays Basic Protocol 5: Testing the capturing abilities of target capturing agents by precipitation/turbidity assays Basic Protocol 6: Capturing of targets (lectins and glycoproteins) in the crude extracts by target capturing agents and separation of the target‐TCA complex from other components of the crude extracts Basic Protocol 7: Releasing the captured targets (lectins and glycoproteins) by dissolving the complex Basic Protocol 8: Separation of the targets (lectins and glycoproteins) from their respective target capturing agents Basic Protocol 9: Verification of the purity of the isolated targets (lectins or glycoproteins)

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A capture and release method based on noncovalent ligand cross-linking and facile filtration for purification of lectins and glycoproteins

Cited by 12 publications

References 45 publications

Electrochemically produced local pH changes stimulating (bio)molecule release from pH-switchable electrode-immobilized avidin–biotin systems

Electrochemically produced local pH changes stimulating (bio)molecule release from pH-switchable electrode-immobilized avidin–biotin systems

A Two-Dimensional Affinity Capture and Separation Mini-Platform for the Isolation, Enrichment, and Quantification of Biomarkers and Its Potential Use for Liquid Biopsy

A Rapid and Facile Purification Method for Glycan‐Binding Proteins and Glycoproteins

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