Provided are in vitro serologic methods of assessing the presence of, and assessing the progression of, liver fibrosis in a subject. Also provided are methods of assessing efficacy of an agent for the treatment of liver fibrosis, and methods of treating liver fibrosis. The methods involve quantitatively measuring di-sialylated and mono-sialylated O-glycoforms of a peptide fragment of hemopexin (HPX) in a test serum sample obtained from a test subject, and comparing the measured amounts of di-sialylated and mono-sialylated O-glycoforms of the peptide fragment of hemopexin to a reference amount. In certain embodiments, the measuring is performed using LC-MS/MS-MRM (liquid chromatography/tandem mass spectrometry/multiple reaction monitoring). In certain embodiments, the measuring is performed using LC/MS3 (liquid chromatography with triple-stage mass spectrometric detection).

The disclosure directed to methods for measuring an analyte alone or in combination with total protein in biological samples. More particularly, the disclosure relates to methods for measuring an analyte and/or total protein using one or more colorimetric reagents alone or in combination with protein precipitation reagents.

The present invention provides a method, wherein the method classifies a subject as suffering from dry eye, the method consisting of: a. obtaining demographic data, consisting of the age and gender of the subject; b. obtaining a tear sample from the patient, and determining the level of human serum albumin; c. from the determined level of human serum albumin, assigning a score for the determined amount of human serum albumin; and d. from the assigned score, calculating a cutoff probability score, according to the following equation: wherein the subject has dry eye, if the calculated cutoff probability score is from 50% to 60%.

[00001]$\frac{\exp \left(-0.6491-1.1142*\mathrm{Albumin}\right)}{1+\exp \left(-0.6491-1.1142*\mathrm{Albumin}\right)}$

This present disclosure provides methods and compositions that can be used to quantify cfDNA in biofluids using a hybridization approach.

Disclosed is a method comprising acquiring a value relating to VEGF-A of a subject, wherein the value is a measured value of total VEGF-A in a blood sample, or a value obtained by dividing a measured value of VEGF-A.sub.165b in the blood sample by a measured value of total VEGF-A in the blood sample (VEGF-A.sub.165b/total VEGF-A), and the value suggests prognosis of myocardial infarction of the subject or severity of coronary artery disease of the subject.

There is disclosed herein a method of quantitatively determining the concentration of at least one analyte in a sample, the method comprising the steps of either: (i) adding a portion of the sample to a first analyte assay formulation and to an analyte assay reference formulation to generate a first analyte sample and analyte reference sample and determining the concentration of the at least one analyte in the sample and/or (ii) adding a portion of the sample to a second analyte assay formulation and determining the concentration of the at least one analyte in the sample. Also disclosed herein are formulation, kits of parts, systems and computer implemented methods associated with said method.

There is disclosed herein a method of quantitatively determining the concentration of at least one analyte in a sample, the method comprising the steps of either: (i) adding a portion of the sample to a first analyte assay formulation and to an analyte assay reference formulation to generate a first analyte sample and analyte reference sample and determining the concentration of the at least one analyte in the sample and/or (ii) adding a portion of the sample to a second analyte assay formulation and determining the concentration of the at least one analyte in the sample. Also disclosed herein are formulation, kits of parts, systems and computer implemented methods associated with said method.

Disclosed herein are methods of protein quantification and normalization using haloalkylated tryptophan fluorescence. Complex protein samples, i.e., samples that each contain 1,000 or more distinct proteins, from diverse sources that do not have common protein profiles are treated with a halo-substituted organic compound (i.e. haloalkane) that reacts with tryptophan residues to form fluorescent products. Irradiation of the samples with ultraviolet light and the detection and quantification of the resultant fluorescent emissions from all proteins in each sample are then used to obtain comparative values for total protein content among the various samples. The values thus obtained are found to be valid indications of comparative total protein content, despite the fact that the tryptophan levels vary widely among the various proteins in any single sample and the samples, due to the diversity of their origins, tend to differ among themselves in the identities and relative amounts of the proteins that they contain. Protein samples are also normalized to correct for differences in sample dilution, sample loading, and protein transfer inconsistencies, by using stain-free detection of total protein in each of the samples, or detection of subsamples within each sample.

An object of the present invention is to provide a method of measuring a glycated-albumin (GA) value traceable to a certified GA value conveniently at a low cost in a short period of time. A method of measuring a GA value traceable to the certified GA value includes the following steps: A.sub.1) determining Regression equation III by steps including the following b), c), and e.sub.0) to h); b) determining a GA concentration [.sub.L] (A) of two or more GA certified reference materials having respectively different certified GA values; c) measuring a GA-derived absorbance (D) of the two or more GA certified reference materials; e.sub.0) creating Linear equation II.sub.0 having an intercept of zero; f) determining two or more GA concentrations.sub.[P] (E) proportionate to the GA-derived absorbance (D) according to Equation II.sub.0; g) determining two or more GA values (F); and h) creating Regression equation III.

The disclosure provides methods of forming one or more single-cell proteomic samples, such as by: dispensing n droplets of lysis buffer onto a substantially planar solid surface, wherein n>2: dispensing a single cell into each of the n droplets of lysis buffer to produce n droplets with a lysed single cell: dispensing digestion buffer into each of the n droplets to digest proteins from each lysed single cell to produce n droplets comprising peptides: dispensing a chemical tag into at least a subset of the n droplets comprising the peptides to produce labeled peptides, thereby enabling the labeled peptides in a given droplet to be distinguishable from labeled peptides in at least one other droplet: and applying a fluid to merge at least a subset of the droplets into a combined droplet on the substantially planar surface, thereby combining the labeled peptides to form a single-cell proteomic sample.