The purification and identification of human blood serum proteins with affinity to the antitumor active RL2 lactaptin using magnetic microparticles

Abstract The thiol groups of human blood serum proteins were determined after 24 hours interaction with dithionitrobenzoicacid (DTNB) to an average of 538 ± 60 µmol/l serum. After treatment of the serum with [35S]DTNB , autoradiograms of the protein elpherograms revealed two main peaks: The first with 63% of total activity, in the albumin region, corresponding to 0.60 SH/mol, the second with 23% of total activity, in the 7-globulin range, corresponding to 2.2 SH/mol. After 30 minutes incubation with D TNB , or with p-chloromercuribenzoate (CMB), in freshly prepared pools of IgG only 0.2 SH/mol were found which is the expected value already known from the literature.Autoradiograms taken from serum protein elpherograms after interaction with [UC] CMB only show the main SH-peak in the albumin range. Thus ist is concluded that the SH-peak in the γ-globulin region after 24 hours incubation with [35S]DTNB is due to one highly labile S-S-bond which easily undergoes a disulfide exchange with DTNB .

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In vitro Distribution of Americium in Human Blood Serum Proteins

Nature ◽

10.1038/2101283a0 ◽

1966 ◽

Vol 210 (5042) ◽

pp. 1283-1284 ◽

Cited By ~ 10

Author(s):

G. BOOCOCK ◽

D. S. POPPLEWELL

Keyword(s):

Blood Serum ◽

Human Blood ◽

Serum Proteins ◽

Human Blood Serum ◽

Blood Serum Proteins

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Polymeric Affinity Type Adsorbents in the Study of Physiologically Active Substances. Part XX. Synthesis and Use of Iodine Derivatives of Phenolphthalein and Tyrosine in Affinity Chromatography of Human Blood Serum Proteins

Pharmaceutical Chemistry Journal ◽

10.1023/b:phac.0000022086.64640.f0 ◽

2003 ◽

Vol 37 (12) ◽

pp. 663-666

Author(s):

E. G. Polenok ◽

P. V. Kuznetsov

Keyword(s):

Affinity Chromatography ◽

Blood Serum ◽

Human Blood ◽

Serum Proteins ◽

Human Blood Serum ◽

Blood Serum Proteins ◽

Physiologically Active Substances ◽

Derivatives Of ◽

Active Substances

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Influence of liposome charge and composition on their interaction with human blood serum proteins

Molecular and Cellular Biochemistry ◽

10.1007/bf00926084 ◽

1993 ◽

Vol 120 (2) ◽

pp. 119-126 ◽

Cited By ~ 64

Author(s):

Trinidad Hern�ndez-Caselles ◽

Jos� Villala�n ◽

Juan C. G�mez-Fern�ndez

Keyword(s):

Blood Serum ◽

Human Blood ◽

Serum Proteins ◽

Human Blood Serum ◽

Blood Serum Proteins

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Quench me if you can: Alpha-2-macroglobulin trypsin complexes enable serum biomarker analysis by MALDI mass spectrometry

10.1101/2020.11.23.394759 ◽

2020 ◽

Author(s):

Aleksandr S. Taraskin ◽

Konstantin K. Semenov ◽

Aleksandr V. Protasov ◽

Alexey A. Lozhkov ◽

Alexandr A. Tyulin ◽

...

Keyword(s):

Mass Spectrometry ◽

Blood Serum ◽

Proteolytic Activity ◽

Human Blood ◽

Serum Proteins ◽

Maldi Mass Spectrometry ◽

Human Blood Serum ◽

Serum Biomarker ◽

Biomarker Analysis

ABSTRACTOne of the main functions of alpha-2-macroglobulin (A2M) in human blood serum is the binding of all classes of protease. It is known that trypsin, after such interaction, possesses modified proteolytic activity. Trypsin first hydrolyzes two bonds in A2M’s ‘bait region’, and the peptide 705VGFYESDVMGR715 is released from A2M. In this work, specifics of the A2M-trypsin interaction were used to determine A2M concentration directly in human blood serum using MALDI mass-spectrometry. Following exogenous addition of trypsin to human blood serum in vitro, the concentration of the VGFYESDVMGR peptide was measured, using its isotopically-labeled analogue (18O), and A2M concentration was calculated. The optimized mass spectrometric approach was verified using a standard method for A2M concentration determination (ELISA) and the relevant statistical analysis methods. It was also shown that trypsin’s modified proteolytic activity in the presence of serum A2M can be used to analyze other serum proteins, including potential biomarkers of pathological processes. Thus, this work describes a promising approach to serum biomarker analysis that can be technically extended in several useful directions.

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