Albumin binds low–molecular-weight molecules, including proteins and peptides, which then acquire its longer half-life, thereby protecting the bound species from kidney clearance. We developed an experimental method to isolate albumin in its native state and to then identify [mass spectrometry (MS) sequencing] the corresponding bound low–molecular-weight molecules. We used this method to analyze pooled sera from a human disease study set (high-risk persons without cancer, n= 40; stage I ovarian cancer, n = 30; stage III ovarian cancer, n = 40) to demonstrate the feasibility of this approach as a discovery method.

Methods

Albumin was isolated by solid-phase affinity capture under native binding and washing conditions. Captured albumin-associated proteins and peptides were separated by gel electrophoresis and subjected to iterative MS sequencing by microcapillary reversed-phase tandem MS. Selected albumin-bound protein fragments were confirmed in human sera by Western blotting and immunocompetition.

Results

In total, 1208 individual protein sequences were predicted from all 3 pools. The predicted sequences were largely fragments derived from proteins with diverse biological functions. More than one third of these fragments were identified by multiple peptide sequences, and more than one half of the identified species were in vivo cleavage products of parent proteins. An estimated 700 serum peptides or proteins were predicted that had not been reported in previous serum databases. Several proteolytic fragments of larger molecules that may be cancer-related were confirmed immunologically in blood by Western blotting and peptide immunocompetition. BRCA2, a 390-kDa low-abundance nuclear protein linked to cancer susceptibility, was represented in sera as a series of specific fragments bound to albumin.

Conclusion

Carrier-protein harvesting provides a rich source of candidate peptides and proteins with potential diverse tissue and cellular origins that may reflect important disease-related information.

Analysis of albumin-associated peptides and proteins from ovarian cancer patients

https://www.journal-of-hepatology.eu/article/S0168-8278(13)00211-0/pdf

Lysophosphatidylcholine acyltransferase 1 altered phospholipid composition and regulated hepatoma progression

MALDI imaging mass spectrometry in cancer research: combining proteomic profiling and histological evaluation.

Measurement of urinary protein is an essential part of the evaluation of chronic kidney disease; it has both diagnostic and prognostic significance. Proteinuria is an independent risk factor for progression of renal disease, but is also independently associated with increased cardiovascular mortality. Despite its far-reaching implications, the definition, diagnosis and treatment of proteinuria can cause confusion in primary care. Early detection of proteinuria in the context of diabetes or otherwise is vital given the potential for intervention to reduce urinary protein losses and improve renal and cardiovascular outcomes. This article will focus on the definition, potential causes and management of proteinuria, including which individuals should be referred to secondary care.

Proteinuria

In the current context of international conflicts and localized terrorist actions, there is unfortunately a permanent threat of attacks with unconventional warfare agents. Among these, biological agents such as toxins, microorganisms, and viruses deserve particular attention owing to their ease of production and dissemination. Mass spectrometry (MS)-based techniques for the detection and quantification of biological agents have a decisive role to play for countermeasures in a scenario of biological attacks. The application of MS to every field of both organic and macromolecular species has in recent years been revolutionized by the development of soft ionization techniques (MALDI and ESI), and by the continuous development of MS technologies (high resolution, accurate mass HR/AM instruments, novel analyzers, hybrid configurations). New possibilities have emerged for exquisite specific and sensitive detection of biological warfare agents. MS-based strategies for clinical application can now address a wide range of analytical questions mainly including issues related to the complexity of biological samples and their available volume. Multiplexed toxin detection, discovery of new markers through omics approaches, and identification of untargeted microbiological or of novel molecular targets are examples of applications. In this paper, we will present these technological advances along with the novel perspectives offered by omics approaches to clinical detection and follow-up.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/jms.3747

Mass spectrometry for the detection of bioterrorism agents: from environmental to clinical applications.

Mass spectrometry-based plasma peptide profiling of acute exacerbation in HBeAg-positive chronic hepatitis B.

Diabetic retinopathy (DR) is a chronic retinal disorder, in which the retinal microvasculature is gradually altered, ultimately leading to blindness. Previous observations on clinical variations of the onset and severity of DR in various patients and populations suggest that genetic polymorphisms contribute to DR development. The present study was undertaken in an attempt to uncover new genetic factors contributing to the development of DR in a Taiwanese population. A well-defined Taiwanese population comprising persons with type 2 diabetes mellitus (T2DM) (n = 749) was recruited for this study. We conducted a genome-wide association study in an independent set of 174 patients with DR and 575 without DR, using Illumina HumanHap550-Duo BeadChip. Eleven single nucleotide polymorphisms (SNPs) with the most significant test statistics (p 6 1 10 5 ) were selected from one of the models. Of the selected SNPs, rs832882 (G/A) (p = 2.29 10 6 ; odds ratio (OR) = 1.49; 95% confidence interval (CI) = 1.11-2.00) and rs3742872 (G/A) (P = 1.19 10 15 ; OR = 1.95; 95% CI = 1.02-3.72), both identified as having the most significant association with DR, are located in the intronic region of the gene encoding the pleckstrin homology (PH) domain-containing proteins family O member 2 (PLEKHO2) and family H member 1 (PLEKHH1), respectively. Functional prediction analysis strengthened the likelihood of participation of PLEKHO2 and PLEKHH1 in the development of DR. The current findings suggest that the rs832882 and rs3742872 polymorphisms may be harbouring retinopathy susceptibility in a Taiwanese population, and implicate the pathological role of PH domain-containing proteins in DR development.

/pdf/association-of-the-plekho2-and-plekhh1-gene-polymorphisms-3xcz2ynt3x.pdf

Association of the PLEKHO2 and PLEKHH1 gene polymorphisms with type 2 diabetic retinopathy in a Taiwanese population

Direct tissue analysis by MALDI-TOF mass spectrometry in human hepatocellular carcinoma

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Papers

Mass spectrometry-based plasma peptide profiling of acute exacerbation in HBeAg-positive chronic hepatitis B.

Association of the PLEKHO2 and PLEKHH1 gene polymorphisms with type 2 diabetic retinopathy in a Taiwanese population

Direct tissue analysis by MALDI-TOF mass spectrometry in human hepatocellular carcinoma