Multiplex Immunochips for High-Accuracy Detection of AFP-L3% Based on Surface-Enhanced Raman Scattering: Implications for Early Liver Cancer Diagnosis

Abstract

α-Fetoprotein (AFP) is an important tumor biomarker. In particular, the overexpression of AFP-L3 is associated with hepatocellular carcinoma (HCC). Accordingly, several hospitals have begun to employ the ratio of AFP-L3 to the total AFP level (AFP-L3%) as new diagnostic evidence for HCC owing to its high diagnostic accuracy. However, current methods of detection for AFP and AFP-L3 are time-consuming, require multiple samples, and lack in sensitivity and specificity. Herein, we present a novel concept for the early diagnosis of HCC based on the combination of Raman frequency shift and intensity change, and developed surface-enhanced Raman scattering (SERS)-based immunochips via AFP-L3%. In the first step of the study, the frequency shift of 4-mercaptobenzoic acid (MBA) was applied for the quantitative determination of total AFP based on the AFP and anti-AFP interaction on MBA-modified silver chips. 5,5-Dithiobis(succinimidyl-2-nitrobenzoate) (DSNB)-modified immunogold was then incorporated with AFP-L3 antibodies for sandwich immunoreaction on the chips. As a result, we found that a typical Raman band intensity of DSNB presented an exponential linear relationship with the concentration of AFP-L3. Thus, the AFP-L3% can be calculated according to the concentrations of AFP-L3 and total AFP. The most important advantage of the proposed method is the combination of AFP-L3% and frequency shifts of SERS, which exhibits excellent reproducibility and high accuracy, and significantly simplifies the conventional detection procedure of AFP-L3%. Application of the proposed method with the serum of patients with HCC demonstrated its great potential in early liver cancer diagnosis.

Decreased IL‑37 expression in hepatocellular carcinoma tissues and liver cancer cell lines

“…Notably, the present study revealed a negative association between serum AFP and IL-37 of cancer tissues, but not in PT. AFP usually increases with the deterioration of hepatocellular carcinogenesis and is a relatively specific clinical index for the diagnosis of primary liver cancer, but the specificity and sensitivity of diagnosis are relatively low (26). Some researchers have tried to combine AFP detection with some interleukin detection for the diagnosis and prognosis of liver cancer (27).…”

New nomogram predicts the recurrence of hepatocellular carcinoma in patients with negative preoperative serum AFP subjected to curative resection

“…Siripongsakun et al and Zhang et al suggested that the postoperative measurement of AFP in these HCC populations may not be helpful and should be omitted because of its poor sensitivity. Although several markers may be used in the prediction and diagnosis of HCC, including AFP‐L3, glypican‐3, cyclase‐associated protein 2 (CAP2), AFP‐mRNA, and the half‐life of serum AFP, there are many limitations to the use of these markers in clinical practice . Many staging systems have been used for the classification and prognostication of HCC, including the TNM system, Okuda score, Japan Integrated Staging score, CLIP score, and BCLC stage .…”

“…Although several markers may be used in the prediction and diagnosis of HCC, including AFP-L3, glypican-3, cyclase-associated protein 2 (CAP2), AFP-mRNA, and the half-life of serum AFP, there are many limitations to the use of these markers in clinical practice. [11][12][13][14] Many staging systems have been used for the classification and prognostication of HCC, including the TNM system, Okuda score, Japan Integrated Staging score, CLIP score, and BCLC stage. [15][16][17][18] Nonetheless, these classification criteria vary considerably, present controversial results, and are divided into different groups with varying prognostic results and therapeutic strategies.…”

Advances in surface‐enhanced Raman spectroscopy for cancer diagnosis and staging

“…Multiplexed detection of low concentration (~5 pM) of AFP and glypican‐3 biomarkers in serum samples of HCC patients have been reported using this method . Quantitative assessment of AFP‐L3 for early diagnosis of HCC has been demonstrated by incorporating sandwich complex and shift immunoassay . Scheme for this study is shown in Figure d.…”