scholarly journals Enzyme Stability in Nanoparticle Preparations Part 1: Bovine Serum Albumin Improves Enzyme Function

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4593
Author(s):  
Jason Thomas Duskey ◽  
Federica da Ros ◽  
Ilaria Ottonelli ◽  
Barbara Zambelli ◽  
Maria Angela Vandelli ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Specific Activity ◽  
Enzyme Stability ◽  
Polymeric Nanoparticles ◽  
Release Kinetics ◽  
Titration Calorimetry ◽  
Therapeutic Effects

Enzymes have gained attention for their role in numerous disease states, calling for research for their efficient delivery. Loading enzymes into polymeric nanoparticles to improve biodistribution, stability, and targeting in vivo has led the field with promising results, but these enzymes still suffer from a degradation effect during the formulation process that leads to lower kinetics and specific activity leading to a loss of therapeutic potential. Stabilizers, such as bovine serum albumin (BSA), can be beneficial, but the knowledge and understanding of their interaction with enzymes are not fully elucidated. To this end, the interaction of BSA with a model enzyme B-Glu, part of the hydrolase class and linked to Gaucher disease, was analyzed. To quantify the natural interaction of beta-glucosidase (B-Glu,) and BSA in solution, isothermal titration calorimetry (ITC) analysis was performed. Afterwards, polymeric nanoparticles encapsulating these complexes were fully characterized, and the encapsulation efficiency, activity of the encapsulated enzyme, and release kinetics of the enzyme were compared. ITC results showed that a natural binding of 1:1 was seen between B-Glu and BSA. Complex concentrations did not affect nanoparticle characteristics which maintained a size between 250 and 350 nm, but increased loading capacity (from 6% to 30%), enzyme activity, and extended-release kinetics (from less than one day to six days) were observed for particles containing higher B-Glu:BSA ratios. These results highlight the importance of understanding enzyme:stabilizer interactions in various nanoparticle systems to improve not only enzyme activity but also biodistribution and release kinetics for improved therapeutic effects. These results will be critical to fully characterize and compare the effect of stabilizers, such as BSA with other, more relevant therapeutic enzymes for central nervous system (CNS) disease treatments.

Interlaboratory Study of the IFCC Method for Alanine Aminotransferase Performed with Use of a Partly Purified Reference Material

1992 ◽  
Vol 38 (12) ◽  
pp. 2365-2371 ◽  
Author(s):  
F Schiele ◽  
J Muller ◽  
E Colinet ◽  
G Siest ◽  
P Arzoglou ◽  
...  
Keyword(s):  
Reference Material ◽  
Bovine Serum Albumin ◽  
Confidence Interval ◽  
Serum Albumin ◽  
Enzyme Activities ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Bovine Serum ◽  
Specific Activity ◽  
Interlaboratory Trial

Abstract We present the results of a study on performance of a reference material for alanine aminotransferase (ALT, EC 2.6.1.2) and the corresponding IFCC-approved method in an interlaboratory trial involving 13 laboratories. The ALT material was partly purified from pig heart (specific activity, 150 kU/g) and was essentially free of six potentially contaminating enzyme activities, including aspartate aminotransferase (EC 2.6.1.1). The partly purified ALT was lyophilized in a triethanolamine-buffered matrix, pH 6.4, containing bovine serum albumin and saccharose. Under these conditions, the predicted yearly loss of activity was 0.02% at 4 degrees C and < 0.01% at -20 degrees C. The final blank-corrected results of the accepted set of data gave a mean (SD) of 128.5 (5.1) U/L. The among-laboratory SD was 4.6 U/L and the within-laboratory SD was 2.0 U/L. The certified ALT catalytic concentration in the reconstituted material was 129 U/L with a 0.95 confidence interval of +/- 4 U/L.

The complexity of condensed tannin binding to bovine serum albumin – An isothermal titration calorimetry study

2016 ◽  
Vol 190 ◽  
pp. 173-178 ◽  
Author(s):  
Rachel L. Kilmister ◽  
Peta Faulkner ◽  
Mark O. Downey ◽  
Samuel J. Darby ◽  
Robert J. Falconer
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Isothermal Titration Calorimetry ◽  
Bovine Serum ◽  
Condensed Tannin ◽  
Titration Calorimetry

Purification of Progesterone Antiserum for Radioimmunoassay, by Chromatography on QAE-Sephadex A-50

1973 ◽  
Vol 19 (9) ◽  
pp. 1027-1030 ◽  
Author(s):  
Frank H Bodley ◽  
Alcide Chapdelaine ◽  
Kenneth D Roberts
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Column Chromatography ◽  
Bovine Serum ◽  
Specific Activity ◽  
Antibody Preparation ◽  
Covalent Coupling

Abstract Antibodies were generated in sheep after administration of the antigen, progesterone-11α-succinyl-bovine serum albumin. This antiserum was purified 10-fold by treatment with "Aerosil" and column chromatography on "QAE-Sephadex A-50." In addition to an increased specific activity (cpm progesterone bound/g of protein) the purified antibody fraction was less cross-reactive with pregnenolone and desoxycorticosterone than was the crude antiserum. A sensitivity of 10 pg of progesterone was obtained with the purified antibody preparation, compared to 25 pg for the crude antiserum. This method of purification of antisera was devised in order to concentrate the antibody fraction before covalent coupling to insoluble supports such as activated arylamine-glass particles.

scholarly journals Formation of Nanocomplexes between Carboxymethyl Inulin and Bovine Serum Albumin via pH-Induced Electrostatic Interaction

Molecules ◽  
2019 ◽  
Vol 24 (17) ◽  
pp. 3056 ◽  
Author(s):  
Guiying Huang ◽  
Jun Liu ◽  
Weiping Jin ◽  
Zihao Wei ◽  
Chi-Tang Ho ◽  
...  
Keyword(s):  
Sodium Chloride ◽  
Bovine Serum Albumin ◽  
Complex Formation ◽  
Serum Albumin ◽  
Electrostatic Interaction ◽  
Bovine Serum ◽  
Titration Calorimetry ◽  
Food Ingredients ◽  
Enthalpy Changes ◽  
Ft Ir

As a functional polysaccharide, inulin was carboxymethylated and it formed nanocomplexes with bovine serum albumin (BSA). The success of obtaining carboxymethyl inulin (CMI) was confirmed by a combination of Fourier transform Infrared (FT-IR), Raman spectroscopy, gel permeation chromatography (GPC), and titration. The effects of pH and ionic strength on the formation of CMI/BSA nanocomplexes were investigated. Our results showed that the formation of complex coacervate (pHφ1) and dissolution of CMI/BSA insoluble complexes (pHφ2) appeared in pH near 4.85 and 2.00 respectively. FT-IR and Raman data confirmed the existence of electrostatic interaction and hydrogen bonding between CMI and BSA. The isothermal titration calorimetry (ITC) results suggested that the process of complex formation was spontaneous and exothermic. The complexation was dominated by enthalpy changes (∆Η < 0, ∆S < 0) at pH 4.00, while it was contributed by enthalpic and entropic changes (∆Η < 0, ∆S > 0) at pH 2.60. Irregularly shaped insoluble complexes and globular soluble nanocomplexes (about 150 nm) were observed in CMI/BSA complexes at pH 4.00 and 2.60 while using optical microscopy and atomic force microscopy, respectively. The sodium chloride suppression effect on CMI/BSA complexes was confirmed by the decrease of incipient pH for soluble complex formation (or pHc) and pHφ1 under different sodium chloride concentrations. This research presents a new functional system with the potential for delivering bioactive food ingredients.

scholarly journals Binding of an Oligomeric Ellagitannin Series to Bovine Serum Albumin (BSA): Analysis by Isothermal Titration Calorimetry (ITC)

2015 ◽  
Vol 63 (49) ◽  
pp. 10647-10654 ◽  
Author(s):  
Maarit Karonen ◽  
Marianne Oraviita ◽  
Irene Mueller-Harvey ◽  
Juha-Pekka Salminen ◽  
Rebecca J. Green
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Isothermal Titration Calorimetry ◽  
Bovine Serum ◽  
Titration Calorimetry
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