Clinical Assay for AFP-L3 by Using Multiple Reaction Monitoring–Mass Spectrometry for Diagnosing Hepatocellular Carcinoma

Kim, Hyunsoo,Sohn, Areum,Yeo, Injoon,Yu, Su Jong,Yoon, Jung-Hwan,Kim, Youngsoo American Association for Clinical Chemistry, Inc. 2018 Clinical chemistry Vol.64 No.8

<P><B>BACKGROUND:</B></P><P><I>Lens culinaris</I> agglutinin-reactive fraction of α-fetoprotein (AFP-L3) is a serum biomarker for hepatocellular carcinoma (HCC). AFP-L3 is typically measured by liquid-phase binding assay (LiBA). However, LiBA does not always reflect AFP-L3 concentrations because of its low analytical sensitivity. Thus, we aimed to develop an analytically sensitive multiple reaction monitoring–mass spectrometry (MRM-MS) assay to quantify AFP-L3 in serum.</P><P><B>METHODS:</B></P><P>The assay entailed the addition of a stable isotope-labeled internal standard protein analog, the enrichment of AFP using a monoclonal antibody, the fractionation of AFP-L3 using <I>L. culinaris</I> agglutinin lectin, deglycosylation, trypsin digestion, online desalting, and MRM-MS analysis. The performance of the MRM-MS assay was compared with that of LiBA in 400 human serum samples (100 chronic hepatitis, 100 liver cirrhosis, and 200 HCC). Integrated multinational guidelines were followed to validate the assay for clinical implementation.</P><P><B>RESULTS:</B></P><P>The lower limit of quantification of the MRM-MS assay (0.051 ng/mL) for AFP-L3 was less than that of LiBA (0.300 ng/mL). Thus, AFP-L3, which was not observed by LiBA in HCC samples (n = 39), was detected by the MRM-MS assay, improving the clinical value of AFP-L3 as a biomarker by switching to a more analytical sensitive platform. The method was validated, meeting all the criteria in integrated multinational guidelines.</P><P><B>CONCLUSIONS:</B></P><P>Because of the lower incidence of false-negative findings, the MRM-MS assay is more suitable than LiBA for early detection of HCC.</P>

Cost-Effective Automated Preparation of Serum Samples for Reproducible Quantitative Clinical Proteomics

Lee, Jihyeon,Kim, Hyunsoo,Sohn, Areum,Yeo, Injoon,Kim, Youngsoo American Chemical Society 2019 Journal of proteome research Vol.18 No.5

<P>Reproducible sample preparation remains a significant challenge in large-scale clinical research using selected reaction monitoring-mass spectrometry (SRM-MS), which enables a highly sensitive multiplexed assay. Although automated liquid-handling platforms have tremendous potential for addressing this issue, the high cost of their consumables is a drawback that renders routine operation expensive. Here we evaluated the performance of a liquid-handling platform in preparing serum samples compared with a standard experiment while reducing the outlay for consumables, such as tips, wasted reagents, and reagent stock plates. A total of 26 multiplex assays were quantified by SRM-MS using four sets of 24 pooled human serum aliquots; the four sets used a fixed number (1, 4, 8, or 24) of tips to dispense digestion reagents. This study demonstrated that the use of 4 or 8 tips is comparable to 24 tips (standard experiment), as evidenced by their coefficients of variation: 13.5% (for 4 and 8 tips) versus 12.0% (24 tips). Thus we can save 37% of the total experimental cost compared with the standard experiment, maintaining nearly equivalent reproducibility. The routine operation of cost-effective liquid-handling platforms can enable researchers to process large-scale samples with high throughput, adding credibility to their findings by minimizing human error.</P> [FIG OMISSION]</BR>

Targeted Proteomics Predicts a Sustained Complete-Response after Transarterial Chemoembolization and Clinical Outcomes in Patients with Hepatocellular Carcinoma: A Prospective Cohort Study

Yu, Su Jong,Kim, Hyunsoo,Min, Hophil,Sohn, Areum,Cho, Young Youn,Yoo, Jeong-Ju,Lee, Dong Hyeon,Cho, Eun Ju,Lee, Jeong-Hoon,Gim, Jungsoo,Park, Taesung,Kim, Yoon Jun,Kim, Chung Yong,Yoon, Jung-Hwan,Kim, American Chemical Society 2017 Journal of proteome research Vol.16 No.3

<P>This study was aimed to identify blood-based biomarkers to predict a sustained complete response (CR) after transarterial chemoembolization (TACE) using targeted proteomics. Consecutive patients with HCC who had undergone TACE were prospectively enrolled (training (n = 100) and validation set (n = 80)). Serum samples were obtained before and 6 months after TACE. Treatment responses were evaluated using the modified Response Evaluation Criteria in Solid Tumors (mRECIST). In the training set, the MRM-MS assay identified five marker candidate proteins (LRG1, APCS, BCHE, C7, and FCN3). When this five-marker panel was combined with the best-performing clinical variables (tumor number, baseline PIVKA, and baseline AFP), the resulting ensemble model had the highest area under the receiver operating curve (AUROC) value in predicting a sustained CR after TACE in the training and validation sets (0.881 and 0.813, respectively). Furthermore, the ensemble model was an independent predictor of rapid progression (hazard ratio (HR), 2.889; 95% confidence interval (CI), 1.612-5.178; P value < 0.001) and overall an unfavorable survival rate (HR, 1.985; 95% CI, 1.024-3.848; P value = 0.042) in the entire population by multivariate analysis. Targeted proteomics-based ensemble model can predict clinical outcomes after TACE. Therefore, this model can aid in determining the best candidates for TACE and the need for adjuvant therapy.</P>

In vivo study for the hemostatic efficacy and foreign body reaction of a new powder-type polysaccharide hemostatic agent

Yoonhyeong Byun,Eun Jin Kim,Areum Lee,Young-Ah Suh,Hee Ju Sohn,Jung Min Lee,Jae Seung Kang,Yoo Jin Choi,Youngmin Han,Hongbeom Kim,Wooil Kwon,Jin-Young Jang 대한외과학회 2022 Annals of Surgical Treatment and Research(ASRT) Vol.102 No.2

Purpose: Various hemostatic agents have been introduced in therapy as postoperative bleeding is a poor prognostic factor for postoperative outcomes. These products can be divided into those that directly promote the hemostatic cascade and those that physically form a barrier by absorbing blood. The latter, powder-type hemostatic agents have the advantages of being inexpensive and more absorbable with less foreign body reactions (FBRs) and are applicable to a relatively wide area. This study was conducted to verify the safety and efficacy of a newly invented polysaccharide product (OOZFIX, Theracion Biomedical), which improves blood absorption and hemostatic effects. Methods: Two separate animal experiments were performed. The first evaluated FBRs histologically at 3 days, 2 weeks, and 4 weeks, after implantation of OOZFIX in rats, and the second compared hemostatic performance of OOZFIX and Arista AH (Bard) in the porcine liver punch biopsy model. Results: We found minimal FBRs in the 3-day group and no reactions in both the 2-week and 4-week groups after implantation of hemostatic agents. The time to hemostasis of OOZFIX was not significantly different from that of Arista AH (median [interquartile range]: 9 [6–10] minutes vs. 8 [6–10] minutes, respectively; P = 0.522). When comparing the serial bleeding grade tendency, there was no statistical difference between OOZFIX and Arista AH (P = 0.656). Conclusion: OOZFIX caused a minimal FBR that disappeared within 2 weeks in vivo, and its hemostatic performance was comparable with that of an existing agent, Arista AH. Further clinical studies are required in the future.

A Multimarker Panel Predicts Complete Response after Transarterial Chemoembolization in Patients with Hepatocellular Carcinoma

( Su Jong Yu ),( Hyunsoo Kim ),( Hophil Min ),( Areum Sohn ),( Young Youn Cho ),( Jeong-ju Yoo ),( Dong Hyeon Lee ),( Eun Ju Cho ),( Jeong-hoon Lee ),( Jungsoo Gim ),( Taesung Park ),( Yoon Jun Kim ) 대한간학회 2016 춘·추계 학술대회 (KASL) Vol.2016 No.1

Aims: Achievement of a complete response (CR) after transarterial chemoembolization (TACE) is the most robust predictor of favorable outcomes in patients with hepatocellular carcinoma (HCC). The aim of this study was to identify blood-based biomarkers to predict a sustained CR after TACE using targeted proteomics. Methods: Consecutive patients with HCC who had undergone TACE were drawn from our prospective cohort [training set (n=100) and validation set (n=80)]. Serum samples were obtained before and 6 months after TACE. Treatment responses were evaluated using the modified Response Evaluation Criteria in Solid Tumors (mRECIST). Multiple reaction monitoring-mass spectrometry (MRM-MS) was used to measure marker candidate proteins (MCPs) with regard to their association with the recurrence of HCC and a sustained CR after TACE. Results: In the training set, the MRM-MS assay identified 5 MCPs (MRM-MS marker panel). When this 5-marker panel was combined with the best-performing clinical variables (tumor number, baseline PIVKA, and baseline AFP), the resulting ensemble model had the highest area under the receiver operating curve (AUROC) value in predicting a sustained CR after TACE in the training and validation sets (0.881 and 0.813, respectively). Further, the ensemble model remained an independent predictor of rapid progression (hazard ratio, 2.889; 95% confidence interval, 1.612-5.178; P<0.001) in the entire population by multivariate analysis. Conclusions: Our ensemble model before TACE can predict a sustained CR after TACE. Therefore, this model can aid in determining the best candidates for TACE and the need for adjuvant therapy.

IPTV 환경에서의 지불 결제 서비스 연구 및 개발

박세철(Park Se Cheol),손영설(Sohn Young Seol),강아름보라(Kang Areum Bora) 한국통신학회 2008 한국통신학회 학술대회논문집 Vol.2008 No.11

Method Validation by CPTAC Guidelines for Multi-protein Marker Assays Using Multiple Reaction Monitoring-mass Spectrometry

Minsoo Son,Hyunsoo Kim,Injoon Yeo,Yoseop Kim,Areum Sohn,Youngsoo Kim 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.2

Quantifying multiple protein biomarkers in a blood sample at one time has many advantages for diagnosing human diseases. In this study, 34 multiplex assays by multiple reaction monitoring-mass spectrometry (MRM-MS) for serum biomarkers were characterized according to Clinical Proteomic Tumor Analysis Consortium (CPTAC) guidelines. The assays revealed that the median lower limit of quantitation (LLOQ) was 0.37 fmol/μL (16.0 ng/mL) and that the median total coefficient of variation (CV) was 18.2%, 12.2%, and 10.6% in the low-, medium-, and high-quality control (QC) samples. With regard to selectivity, the median mean differences in slope and concentration were 2.1% and 4.3%, respectively. The median values for all CVs and %difference from the nominal concentration for stability were 9.5% and 2.7% in low-QC and 3.8% and 3.1% in medium-QC. The median total CV was 9.8% in the reproducibility. Finally, 17 protein-based biomarker assays were reliable and transferrable for preclinical purposes per CPTAC guidelines.