Ensuring sample quality for blood biomarker studies in clinical trials

A multicenter international study for plasma and serum sample preparation

Feng Ming Kong, Lujun Zhao, Luhua Wang, Yuhchyau Chen, Jie Hu, Xiaolong Fu, Chunxue Bai, Li Wang, Theodore S. Lawrence, Mitchell S. Anscher, Adam Dicker, Paul Okunieff

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.

Original languageEnglish (US)
Pages (from-to)625-634
Number of pages10
JournalTranslational Lung Cancer Research
Volume6
Issue number6
DOIs
StatePublished - Dec 1 2017

Fingerprint

Multicenter Studies
Biomarkers
Platelet Factor 4
Clinical Trials
Serum
Blood Platelets
Ice
Asian Americans
Centrifugation
Healthy Volunteers
Enzyme-Linked Immunosorbent Assay
Temperature

Keywords

  • Biomarker study
  • Plasma
  • Platelet factor 4 (PF4)
  • Serum

ASJC Scopus subject areas

  • Oncology

Cite this

Ensuring sample quality for blood biomarker studies in clinical trials : A multicenter international study for plasma and serum sample preparation. / Kong, Feng Ming; Zhao, Lujun; Wang, Luhua; Chen, Yuhchyau; Hu, Jie; Fu, Xiaolong; Bai, Chunxue; Wang, Li; Lawrence, Theodore S.; Anscher, Mitchell S.; Dicker, Adam; Okunieff, Paul.

In: Translational Lung Cancer Research, Vol. 6, No. 6, 01.12.2017, p. 625-634.

Research output: Contribution to journalArticle

Kong, FM, Zhao, L, Wang, L, Chen, Y, Hu, J, Fu, X, Bai, C, Wang, L, Lawrence, TS, Anscher, MS, Dicker, A & Okunieff, P 2017, 'Ensuring sample quality for blood biomarker studies in clinical trials: A multicenter international study for plasma and serum sample preparation', Translational Lung Cancer Research, vol. 6, no. 6, pp. 625-634. https://doi.org/10.21037/tlcr.2017.09.13
Kong, Feng Ming ; Zhao, Lujun ; Wang, Luhua ; Chen, Yuhchyau ; Hu, Jie ; Fu, Xiaolong ; Bai, Chunxue ; Wang, Li ; Lawrence, Theodore S. ; Anscher, Mitchell S. ; Dicker, Adam ; Okunieff, Paul. / Ensuring sample quality for blood biomarker studies in clinical trials : A multicenter international study for plasma and serum sample preparation. In: Translational Lung Cancer Research. 2017 ; Vol. 6, No. 6. pp. 625-634.
@article{c6ba5305fc95439f87151e9057017def,
title = "Ensuring sample quality for blood biomarker studies in clinical trials: A multicenter international study for plasma and serum sample preparation",
abstract = "Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.",
keywords = "Biomarker study, Plasma, Platelet factor 4 (PF4), Serum",
author = "Kong, {Feng Ming} and Lujun Zhao and Luhua Wang and Yuhchyau Chen and Jie Hu and Xiaolong Fu and Chunxue Bai and Li Wang and Lawrence, {Theodore S.} and Anscher, {Mitchell S.} and Adam Dicker and Paul Okunieff",
year = "2017",
month = "12",
day = "1",
doi = "10.21037/tlcr.2017.09.13",
language = "English (US)",
volume = "6",
pages = "625--634",
journal = "Translational Lung Cancer Research",
issn = "2226-4477",
publisher = "Society for Translational Medicine (STM)",
number = "6",

}

TY - JOUR

T1 - Ensuring sample quality for blood biomarker studies in clinical trials

T2 - A multicenter international study for plasma and serum sample preparation

AU - Kong, Feng Ming

AU - Zhao, Lujun

AU - Wang, Luhua

AU - Chen, Yuhchyau

AU - Hu, Jie

AU - Fu, Xiaolong

AU - Bai, Chunxue

AU - Wang, Li

AU - Lawrence, Theodore S.

AU - Anscher, Mitchell S.

AU - Dicker, Adam

AU - Okunieff, Paul

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.

AB - Background: Sample quality is critical for biomarker detection in oncology, and platelet degradation and contamination in plasma have a remarkable impact on the ability to accurately quantify many blood-based biomarkers. Platelet factor 4 (PF4) can be used as an indicator to monitor sample quality. This multicenter study aimed to determine the impact of critical components of the blood sample handling process on platelet degradation/contamination and to establish an optimal method for collecting platelet-poor plasma samples. Methods: At each of six participating centers, blood samples were drawn from 12-13 healthy volunteers. Serum and plasma samples were prepared from whole blood samples using nine different methods that have been commonly used in ongoing multicenter trials. PF4 levels in the prepared samples were measured by enzyme-linked immunosorbent assay (ELISA). Paired t-tests were used for statistical analysis. Results: Blood samples were collected from 74 subjects enrolled in six centers. PF4 levels were significantly higher in serum samples than in plasma samples (P < 0.001), in plasma samples from blood that sat at room temperature for 5 minutes (P=0.021), in plasma samples prepared at an insufficient centrifugal force (P < 0.001), and in plasma samples prepared from blood that sat for longer than 4 hours on ice (P=0.001). For each method, the PF4 levels did not differ significantly among the centers or between Chinese and American subjects. The methods that resulted in normal levels of PF4 involved keeping blood samples on ice for 30 minutes to < 4 hours and centrifugation at 2,500-3,000 ×g for 30 min. Conclusions: This multicenter study evaluated multiple blood sample handling conditions for minimizing platelet degradation during plasma serum preparation and determined an optimal method for preparing platelet-poor plasma. The findings of this study can be applied in future blood biomarker studies.

KW - Biomarker study

KW - Plasma

KW - Platelet factor 4 (PF4)

KW - Serum

UR - http://www.scopus.com/inward/record.url?scp=85031323584&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85031323584&partnerID=8YFLogxK

U2 - 10.21037/tlcr.2017.09.13

DO - 10.21037/tlcr.2017.09.13

M3 - Article

VL - 6

SP - 625

EP - 634

JO - Translational Lung Cancer Research

JF - Translational Lung Cancer Research

SN - 2226-4477

IS - 6

ER -