Selenium binding protein 1 detection from body fluids for diagnosis of peracute tissue damage
A tissue injury and acute technique, applied in disease diagnosis, microbial determination/inspection, biological testing, etc., can solve problems such as incomplete specificity of cardiomyocytes
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Embodiment 1
[0124] Example 1: SELENBP1 can be detected in blood samples from patients with acute coronary syndrome
[0125] Circulating SELENBP1 was determined in serum samples from patients suspected of having acute coronary syndrome (ACS). Diagnosis was confirmed in parallel by traditional methods. Blood samples were drawn at several time points after the first onset of symptoms suspected to be caused by ACS. Patients had higher mean SELENBP1 concentrations than healthy controls. Concentrations display a dynamic time course with patient-specific features.
Embodiment 2
[0126] Example 2: SELENBP1 in blood of healthy controls
[0127] High quality serum samples from a panel of control adults (n=75) were used to determine normal SELENBP1 levels in the blood of healthy subjects. Only trace concentrations were detectable, mainly below the linear range of the SELENBP1 assay. The values obtained were normally distributed with a mean value of 0.23 nmol / l. The 95th percentile of values for SELENBP1 concentrations in healthy controls was up to 0.32 nmol / l and the 99th percentile was up to 0.39 nmol / l.
Embodiment 3
[0128] Example 3: Expression and purification of recombinant SELENBP1
[0129]Expression of recombinant human SELENBP1 (rhSELENBP1 ) in baculovirus-infected insect cells. Using primers P1 (SEQ: ID NO.2) and P2 (SEQ: ID NO.3) (Invitrogen, Thermo Fischer Scientific, Dreieich, Germany) containing BamHI and HindIII restriction sites, respectively, according to SEQ: ID NO.1 The cDNA sequence encoding rhSELENBP1 (according to UniProtKB database No. Q13228-1) was amplified by PCR. The pFastBac1 plasmid (Thermo Fischer Scientific) was digested with BamHI and HindIII, and the plasmid fragment was removed and replaced by the PCR sequence resulting in the pFastBac2-SelenBP1-His6 plasmid. Transform DH10Bac E. coli cells and identify bacmid positive cells, culture and isolate recombinant bacmid. Sf9 insect cells were transferred with bacmid DNA by Cellfectin (Thermo Fischer Scientific) for obtaining recombinant virus stock for initiating rhSELENBP1 biosynthesis in "High Five" insect susp...
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