Disclosed herein are methods for making a transcriptome-wide expression profile of a biological sample and identifying biomarkers that can be used to diagnose, monitor the onset, monitor the progression, and assess the recovery of a disease in a subject. The biomarkers can also be used to establish and evaluate treatment regimens.

Provided herein are compositions, systems, and methods for monitoring platelet activation. In particular, provided herein are sensor devices for measuring platelet activation and uses thereof.

The present invention provides a humanized antibody or antibody fragment comprising (a) a humanized light chain comprising 1) Complementarity Determining Region (CDR)-L1, the sequence of which is identical to the sequence of SEQ ID NO: 3; 2) CDR-L2, the sequence of which is identical to the sequence of SEQ ID NO: 4; and 3) CDR-L3, the sequence of which is identical to the sequence of SEQ ID NO: 5, and (b) a humanized heavy chain comprising 1) CDR-H1, the sequence of which is identical to the sequence of SEQ ID NO: 6; 2) CDR-H2, the sequence of which is identical to the sequence of SEQ ID NO: 7; and 3) CDR-H3, the sequence of which is identical to the sequence of SEQ ID NO: 8, as well as methods for treating, diagnosing, and monitoring the progression of HIT. The present invention also provides methods for assessing the antigenicity and ability to cause HIT of anionic anticoagulants. The present invention also provides a mutant protein which has the same amino acid sequence of a wild type PF4 monomer except that (i) at least one amino acid of the wild type PF4 monomer has been deleted, (ii) at least one amino acid of the wild type PF4 monomer has been replaced by another amino acid, or (iii) a combination of such changes has been made. The present invention also provides methods of treating or reducing the likelihood of HIT, treating angiogenesis, treating abnormal cell growth, or affecting coagulation pathologies that lead to thrombus formation, by administering such mutant proteins to a patient.

The present invention provides a method of detecting platelet activation in a patient, the method comprising the steps of a) obtaining a blood sample from a patient suspected of having heparin-induced thrombocytopenia (HIT); b) incubating an effective amount of platelet factor 4 (PF4) with a sample of platelets to yield a sample of PF4-treated platelets; c) contacting the patient blood sample with the PF4-treated platelets; and d) measuring the extent of platelet activation, wherein an increase in platelet activation compared with results obtained using a normal blood sample is indicative of the patient having HIT.

The current disclosure provides antibodies that bind to peptides associated with the primary immunodeficiency disorders (PIDD) Wiskott-Aldrich Syndrome (WAS) and X-linked agammaglobulinemia (XLA). The antibodies can be used in peptide immunoaffinity enrichment coupled to selected reaction monitoring mass spectrometry (immuno-SRM) assays for clinical diagnosis and newborn screening of WAS and XLA, among other uses.

Methods are provided for determining the presence of antibodies in blood or a blood product, using immobilized self-assembled polypeptides comprising an ectodomain and being recognized by the antibodies. The self-assembled polypeptide comprises at least a first chimeric polypeptide. In the methods the functionality and active conformation of the immobilized and self-assembled polypeptides is preserved. Processes for making the immobilized self-assembled polypeptides are also provided.

The present invention relates to a functional, easily automatable assay for establishing a heparin-induced thrombocytopenia (HIT). What is measured is the secretion of PF4 (platelet factor 4) from activated thrombocytes.

A system and method are provided for harvesting target biological substances. The system includes a substrate and a first and second channel formed in the substrate. The channels longitudinally extending substantially parallel to each other. A series of gaps extend from the first channel to the second channel to create a fluid communication path passing between a series of columns with the columns being longitudinally separated by a predetermined separation distance. The system also includes a first source configured to selectively introduce into the first channel a first biological composition at a first channel flow rate and a second source configured to selectively introduce into the second channel a second biological composition at a second channel flow rate. The sources are configured to create a differential between the first and second channel flow rates to generate physiological shear rates along the second channel that are bounded within a predetermined range.

Deamidation is usually viewed as a post-translational modification that sets an expiration date on proteins. Among apoptosis regulators of the Bcl-2 family, Bcl-xL shows a unique eligibility to be either singly or doubly deamidated. The inventors therefore analysed Bcl-xL deamidation state in platelets from mice models where platelets lifespan was manipulated. In parallel, the inventors compared human platelets obtained at steady state from healthy controls, to platelets newly synthesized after recovery from acute thrombocytopenia: they found that while expression levels of Asn52 monodeamidated Bcl-xL remains unchanged, Asn52Asn66 doubly-deamidated Bcl-xL is virtually absent in young platelets and accumulates in old platelets. Therefor the Asn52Asn66 doubly-deamidated Bcl-xL could be used as a reliable biomarker for determining the age of platelets.

A system and method are provided for harvesting target biological substances. The system includes a substrate and a first and second channel formed in the substrate. The channels longitudinally extending substantially parallel to each other. A series of gaps extend from the first channel to the second channel to create a fluid communication path passing between a series of columns with the columns being longitudinally separated by a predetermined separation distance. The system also includes a first source configured to selectively introduce into the first channel a first biological composition at a first channel flow rate and a second source configured to selectively introduce into the second channel a second biological composition at a second channel flow rate. The sources are configured to create a differential between the first and second channel flow rates to generate physiological shear rates along the second channel that are bounded within a predetermined range.