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Last Updated: 06/19/19

BPV Experimental Design

BBRB's Biospecimen Pre-analytical Variables (BPV) Program was designed to systematically investigate the effects of individual pre-analytical factors on the molecular profiles of biospecimens. At the outset of the BPV program, a Scientific Steering Committee was convened to help the program identify the most important parameters for study. The pre-analytical factors identified as contributing to difficulties in obtaining reproducible research results include key parameters related to a) formalin fixation and paraffin embedding (FFPE) of tissues, and b) freezing of tissues and blood. The pre-analytical factors evaluated in BPV included:

  • The delay time, otherwise known as cold ischemia time, between removal of tissues from the body and placement in formalin for FFPE processing
  • The total time in formalin before further processing of FFPE tissues
  • The method of freezing tumor tissue and the temperature at which tissues are frozen
  • The storage temperature of frozen tumor tissue
  • The storage temperature and time of frozen plasma

The BPV program set up experimental protocols to determine the effects of the preanalytical factors listed above. Highly annotated biospecimens were collected for the BPV program through an organized and well controlled infrastructure. Patients were screened, consented, and enrolled in the BPV study at four Biospecimen Source Sites (BSS): Boston Medical Center, Emory University, the University of New Mexico, and the University of Pittsburg Medical Center. The BSS collected renal cell carcinoma, ovarian, fallopian tube, and primary peritoneal carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, and colorectal adenocarcinoma specimens along with matched blood samples and, when possible, adjacent "normal" tissue.

Protocol Overview

Tumors were surgically removed and dissected into six blocks. One block was frozen in liquid nitrogen (LN2) to be analyzed side by side with FFPE tissues in later molecular analyses. A second block was processed for FFPE and used as a histological control tissue for the analyses. The histology (H&E) section was reviewed by a pathologist at the local BSS and by BPVís Pathology Resource Center (PRC) for many parameters including confirmation of diagnosis and the requirement of at least 50% tumor cells by surface area and less than 20% necrosis present. The four remaining blocks were randomly assigned to an experimental protocol that examined one or more of the key preanalytical factors listed above.

Analytical Platforms

Analytical platforms utilized for the BPV program and additional details about BPV operations can be found here.


The results of this study are being analyzed and used to develop evidence-based best practices for optimal collection, processing, and storage of biospecimens. Moreover, accumulated data from these activities will be widely disseminated to the research community to support the development of international standards for biobanking and utilization of biospecimens in cancer research. Please see BPV Publications for a current list of publications from the BPV program.


A BPV sub-study was conducted to explore the ethical, legal and social implications (ELSI) related to consent for broad future research use of the donated biospecimens. Click here to learn more.

Access to Collected Biospecimens

Remaining biospecimens are maintained at the Van Andel Research Institute (VARI) and available for future research use. Please visit VARIís BPV site for more information.

Contribute to Biospecimen Science

Help contribute to a better understanding of how pre-analytical factors affect molecular profiles and specimen quality. View our collaborative announcement here.

Biospecimen Pre-analytical Variables (BPV)