Standard Operating Procedures for Serum and Plasma Collection: Early Detection Research Network Consensus Statement Standard Operating Procedure Integration Working Group

Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan 48109-0725, USA.
Journal of Proteome Research (Impact Factor: 4.25). 01/2009; 8(1):113-7. DOI: 10.1021/pr800545q
Source: PubMed

ABSTRACT Specimen collection is an integral component of clinical research. Specimens from subjects with various stages of cancers or other conditions, as well as those without disease, are critical tools in the hunt for biomarkers, predictors, or tests that will detect serious diseases earlier or more readily than currently possible. Analytic methodologies evolve quickly. Access to high-quality specimens, collected and handled in standardized ways that minimize potential bias or confounding factors, is key to the "bench to bedside" aim of translational research. It is essential that standard operating procedures, "the how" of creating the repositories, be defined prospectively when designing clinical trials. Small differences in the processing or handling of a specimen can have dramatic effects in analytical reliability and reproducibility, especially when multiplex methods are used. A representative working group, Standard Operating Procedures Internal Working Group (SOPIWG), comprised of members from across Early Detection Research Network (EDRN) was formed to develop standard operating procedures (SOPs) for various types of specimens collected and managed for our biomarker discovery and validation work. This report presents our consensus on SOPs for the collection, processing, handling, and storage of serum and plasma for biomarker discovery and validation.

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    • "Serum samples were collected following uniform processing protocols recommended by the EDRN [49]. All samples were allowed to clot and serum was recovered by centrifugation within 2–8 hours of collection and stored at -80°C. "
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    ABSTRACT: Background CT screening for lung cancer is effective in reducing mortality, but there are areas of concern, including a positive predictive value of 4% and development of interval cancers. A blood test that could manage these limitations would be useful, but development of such tests has been impaired by variations in blood collection that may lead to poor reproducibility across populations. Results Blood-based proteomic profiles were generated with SOMAscan technology, which measured 1033 proteins. First, preanalytic variability was evaluated with Sample Mapping Vectors (SMV), which are panels of proteins that detect confounders in protein levels related to sample collection. A subset of well collected serum samples not influenced by preanalytic variability was selected for discovery of lung cancer biomarkers. The impact of sample collection variation on these candidate markers was tested in the subset of samples with higher SMV scores so that the most robust markers could be used to create disease classifiers. The discovery sample set (n = 363) was from a multi-center study of 94 non-small cell lung cancer (NSCLC) cases and 269 long-term smokers and benign pulmonary nodule controls. The analysis resulted in a 7-marker panel with an AUC of 0.85 for all cases (68% adenocarcinoma, 32% squamous) and an AUC of 0.93 for squamous cell carcinoma in particular. This panel was validated by making blinded predictions in two independent cohorts (n = 138 in the first validation and n = 135 in the second). The model was recalibrated for a panel format prior to unblinding the second cohort. The AUCs overall were 0.81 and 0.77, and for squamous cell tumors alone were 0.89 and 0.87. The estimated negative predictive value for a 15% disease prevalence was 93% overall and 99% for squamous lung tumors. The proteins in the classifier function in destruction of the extracellular matrix, metabolic homeostasis and inflammation. Conclusions Selecting biomarkers resistant to sample processing variation led to robust lung cancer biomarkers that performed consistently in independent validations. They form a sensitive signature for detection of lung cancer, especially squamous cell histology. This non-invasive test could be used to improve the positive predictive value of CT screening, with the potential to avoid invasive evaluation of nonmalignant pulmonary nodules.
    Clinical Proteomics 08/2014; 11(1):32. DOI:10.1186/1559-0275-11-32
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    • "Blood samples from TT immunized and unimmunized animals were collected and the sera was separated using reported standard protocol (Tuck et al., 2009). To this equimolar concentrations of TT and CsAuNPs (TT:CsAuNPs) were incubated at 37 C and subjected to particle size and zeta potential analysis at predetermined intervals (0, 6, 12, 24, 48 h). "
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    ABSTRACT: Many biotherapeutic applications of gold nanoparticles make use of conjugated or adsorbed protein moieties. Physical parameters of association such as particle size, morphology, surface chemistry, temperature influences the protein-nanoparticle association and thereby their interaction with the biological environment. In present study, effect of size of chitosan reduced gold nanoparticles (CsAuNPs) and association temperature on structure and function of tetanus toxoid (TT) vaccine has been investigated. CsAuNPs were synthesized in the sizes of 20±3, 40±5 and 80±7nm followed by loading of TT. Binding process of CsAuNPs with TT was investigated at their predetermined micro molar concentrations. Upon binding of TT onto CsAuNPs, particle surface was characterized using X-ray photoelectron spectroscopy. CD spectroscopic evaluation of TT bound 20nm CsAuNPs led to 75% reduction in secondary structure of TT and thereby compromised immune function. Binding of TT with 40 and 80nm sized CsAuNPs did not cause significant modifications in secondary structure or function of TT. Thermodynamic studies using temperature dependent fluorescence spectroscopy revealed an increase in association constants with the temperature. Based on thermodynamic data three phases in CsAuNPs and TT association process were traced. Samples from these distinct phases were also investigated for immunological recognition. Ex-vivo interaction of TT-CsAuNPs with TT positive and negative sera followed by relative change in particle size and zeta potential was studied. The findings here suggests prominent role of particle size and association temperature on adsorbed TT structure and function. Such studies may help in engineering functional nanotherapeutics.
    International Journal of Pharmaceutics 08/2014; 471(s 1–2):439–448. DOI:10.1016/j.ijpharm.2014.06.002 · 3.65 Impact Factor
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    • "Briefly, it is collected from thousands of healthy donors, randomized, standardized and specially prepared according to Clinical Laboratory Improvement Amendments certification procedure approved by FDA (Tuck et al., 2009). Finally, it is provided in dried lyophilized powder (Tuck et al., 2009). The standardized serum can be used in different clinical research studies (Ipp & Gold, 1988; Uchida et al., 2014). "
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    ABSTRACT: International Journal of Pharmaceutics 473 (2014) 599–606. ABSTRACT: The development of medical nanosystems requires knowledge of their behaviour in vivo. Clinical chemistry tests are widely used to estimate the systemic toxicity of nanoparticles. In this paper we have explored the impact of small positively charged nanoparticles – poly(amidoamine), phosphorous and carbosilane dendrimers – on biochemical parameters of standardized serum in vitro. All the dendrimers could shift the main biochemical parameters. Thus, in the case of patients having the normal, but ‘boundary’, values of biochemical parameters, nanoparticle-induced changes can be wrongly interpreted as evidence of some dysfunctions (hepatic, renal, etc.). Summarizing, the cationic polymeric nanoparticles had 3 different types of effect on clinical chemistry tests. First, they were able to change enzyme activities. Second, they interacted with test reagents (substrates) but not with a protein. Third, they had no effect on some tests. Moreover, the effects of nanoparticles of metals, carbon nanotubes, quantum dots, fullerenes, dendrimers having been sized up to 4000 nm and the hundreds of reactive groups, can be significantly higher. Thus, preliminary evaluation of any nanomaterial in vitro is required in clinical chemistry tests before its application in vivo to draw the correct conclusions and benefit animals
    International Journal of Pharmaceutics 07/2014; 473(1-2). DOI:10.1016/j.ijpharm.2014.07.054 · 3.65 Impact Factor
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