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Discovery of the Long Interspersed Nuclear Element-1 activation product [Open Reading Frame-1 (ORF1) protein] in human blood.

Clinica chimica acta; international journal of clinical chemistry (2018-10-06)
Keivan Hosseinnejad, Tyler Yin, Jeremy T Gaskins, James L Bailen, Saeed A Jortani
ABSTRACT

Long Interspersed Nuclear Element 1 is the only autonomous mobile DNA capable of self-propagation, and is an environmental biomarker that is activated upon an environmental trigger. We have developed an ELISA method to detect and measure Open Reading Frame-1 (ORF1) and have applied it to interrogate serum samples from men with equivocal prostate specific antigen (PSA) results. Polyclonal antibodies were developed using the first 14-amino acid peptide of N-terminal-ORF1 protein. Remnant serum samples from a total of 53 men, ages>50 yr, were analyzed for immunoreactive ORF1 (iORF1) and PSA concentrations; outcomes for the non-biopsied and biopsied groups were also recorded. The dynamic range of the ELISA was between (CV): 2.0 (14%) to 30 ng/mL (1.2%). The total imprecision (within-run/inter-day) was: QC3 = 2.7%/21%, QC6 = 1.1%/18%, and QC20 = 0.33%/11%. The median iORF1 concentration in the non-biopsy group was 14.7 ng/mL (Q1 - Q3: 10.5 - Q3:18.4), which was significantly lower than the Biopsy group at 25.0 ng/mL (Q1 - Q3: 20.0-33.1), P-value = .003. In conclusion, we have developed a competitive ELISA and discovered the presence of iORF1 in serum, which could be used to advance future studies involving ORF1 measurement from blood. In addition, iORF1 may be a complement with the PSA screen to better detect prostate cancer.

MATERIALS
Product Number
Brand
Product Description

Milli-Q
Milli-Q® EQ 7000 ultrapure water purification system, Produces ultrapure (Type 1) water from pure water with a distribution flow rate up to 2 L/min., with system-mounted dispenser
Milli-Q
Milli-Q® EQ 7000 ultrapure water purification system, Produces ultrapure (Type 1) water from pure water with a distribution flow rate up to 2 L/min., with wall-mounted dispenser