Nanobodies’ duo facilitates ultrasensitive serum HER-2/neu immunoassays via enhanced avidity interactions

IF 5.7 2区化学 Q1 CHEMISTRY, ANALYTICAL

Analytica Chimica Acta Pub Date : 2024-11-22 DOI:10.1016/j.aca.2024.343472

Suchanat Boonkaew , Laura Teodori , Mikkel H. Vendelbo , Jørgen Kjems , Elena E. Ferapontova

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引用次数: 0

Abstract

Background

Existing liquid biopsy assays for protein biomarkers of cancer are mostly based on antibodies (Ab) contributing unfavorably to their high cost. Easy to express and modify in vitro, nanobodies may be a cost-effective alternative to Ab.

Results

We show that serum HER-2/neu, a biomarker and target of aggressive HER-2/neu(+) cancers, can be accurately detected in a 1.2 h electrochemical cellulase-linked sandwich nanobody/aptamer assay on magnetic beads. Using a single nanobody receptor, 2Rs15d or 2Rb17c, reduces immunoassay's sensitivity by 35%–26 %. A combination of two nanobodies as a duo-receptor recovers the sensitivity of the enzyme-linked nanobody/aptamer-sorbent assay (ELNASA) to 11.9 ± 2.8 μC fM⁻¹, due to the avidity effects making the nanobodies-duo binding properties comparable to those of Ab. Down to 0.1 fM HER-2/neu was detected by ELNASA in serum samples, with no interference from other blood-circulating proteins. In a 30 healthy-volunteers trial, ELNASA more accurately than optical ELISA assayed serum HER-2/neu.

Significance

ELNASA performance rivals that of ELISA, yet estimated to be at least 200 times cheaper, due to the lower cost of nanobodies production, and may be better suited for routine clinical analysis of HER-2/neu, particularly, in low- and middle-income settings with limited resources. The ELNASA approach is generic and may be adapted for specific and ultrasensitive analysis of other blood-circulating proteins.

Abstract Image

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纳米抗体双元体通过增强的抗体相互作用促进超灵敏血清HER-2/neu免疫测定

背景现有的癌症蛋白质生物标志物液体活检测定大多基于抗体（Ab），这不利于降低其高昂的成本。我们的研究结果表明，血清中的 HER-2/neu 是侵袭性 HER-2/neu(+) 癌症的生物标记物和靶标，可以通过磁珠上的电化学纤维素酶连接夹心纳米抗体/aptamer 分析法在 1.2 小时内准确检测到。使用单一纳米抗体受体（2Rs15d 或 2Rb17c）会使免疫测定的灵敏度降低 35%-26%。两个纳米抗体作为双受体的组合使酶联纳米抗体/aptamer-吸附测定（ELNASA）的灵敏度恢复到 11.9 ± 2.8 μC fM-1，这是因为渴求效应使纳米抗体-双受体的结合特性与抗体相当。ELNASA能在血清样本中检测到低至0.1 fM的HER-2/neu，不受其他血液循环蛋白的干扰。在一项30名健康志愿者参加的试验中，ELNASA比光学ELISA更准确地测定了血清中的HER-2/neu。ELNASA的性能可与ELISA媲美，但由于纳米抗体的生产成本较低，估计至少便宜200倍，可能更适合HER-2/neu的常规临床分析，特别是在资源有限的中低收入地区。ELNASA 方法具有通用性，可用于其他血液循环蛋白的特异性超灵敏分析。

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来源期刊

Analytica Chimica Acta 化学-分析化学

CiteScore

10.40

自引率

6.50%

发文量

1081

审稿时长

38 days

期刊介绍： Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.