Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.
Adv Exp Med Biol. 2018;1104:259-273. doi: 10.1007/978-981-13-2158-0_13.
This chapter introduces two formats using bio-layer interferometry competition assays to determine the solution K values of weak glycan-protein interactions. This approach overcomes the challenge of determining weak interactions while minimizing the amount of reagents required. Accurate solution K values aid in understanding the complex relationships between monomeric versus multimeric interactions and affinity versus avidity. The assays have been applied to a well-studied lectin (Erythrina crista-galli lectin) and influenza hemagglutinin (X-31). The solution K values determined from this approach are in good agreement with previous reported literature values from isothermal titration calorimetry and NMR. Additionally, this approach appears robust and precise.
本章介绍了两种使用生物层干涉竞争测定法来确定弱糖蛋白相互作用的溶液 K 值的格式。这种方法克服了确定弱相互作用的挑战，同时最大限度地减少了所需试剂的数量。准确的溶液 K 值有助于理解单体与多聚体相互作用以及亲和力与亲合力之间的复杂关系。该测定法已应用于一种研究充分的凝集素（鸡冠刺桐凝集素）和流感血凝素（X-31）。从该方法确定的溶液 K 值与之前报道的等温滴定量热法和 NMR 的文献值吻合良好。此外，该方法似乎具有稳健性和精确性。
