scholarly journals New immunoassay systems based on recombinant human lactoferrin

Author(s):  
D. A. Semenov ◽  
I. I. Vashkevich ◽  
O. V. Sviridov

In this work, soluble and solid phase immunoreagents, including recombinant human lactoferrin (rhLF), a complex of rhLF with europium ions, rabbit antiserum to rhLF, anti-rhLF immunoglobulin purified by antigen-affinity chromatography and the conjugates of this immunoglobulin with an Eu3+ chelate or horseradish peroxidase have been obtained by a combination of biochemical and synthetic methods using rhLF as an initial compound. Biospecific interactions of the reagents in four immunochemical systems were assessed by measuring the enzyme activity or time-resolved fluorescence. The study resulted in the development of fast and precise immunoassays for biologically active rhLF in transgenic goat milk and in protein fractions obtained in the course of pure rhLF manufacture, as well as in pharmaceutical preparations and food additives.

Author(s):  
O. Sviridov

Specific immunoreagents for the immunoassay technologies of recombinant human lactoferrin in food and biotechnological products have been obtained and characterized. This work was financially supported by the State program of scientific research «Chemical technologies and materials» for 2016 – 2020 years.


1992 ◽  
Vol 10 (7) ◽  
pp. 784-789 ◽  
Author(s):  
Pauline P. Ward ◽  
Jing-Y. Lo ◽  
Mairead Duke ◽  
Gregory S. May ◽  
Denis R. Headon ◽  
...  

Author(s):  
D. Bogdanovich ◽  
Е. Petrushko ◽  
V. Kuzniatsova

The concentration of recombinant human lactoferrin (rhLF) in the milk of 22 transgenic goats of different ages was studied: the average concentration of rhLF in the milk of goats-producers of the 4th year of lactation was 13.7% higher than that of goats of the 3rd year of lactation (6.07 and 5.24 g/l, respectively). It was found that the maximum level of recombinant glycoprotein was produced by the mammary gland of animals in the 2nd and 4th quarters of the studied year period. Protein’s concentration in milk measured in the 2nd and 4th quarters was 6.4 g/l and 6.33 g/l for producers of the third year of lactation and 8.14 g/l and 7.02 g/l for goats of the fourth year of lactation, respectively. The correlation between the season of the year and the content of the target protein in milk was detected (p <0.01), it was found that the most active rhLF production in goat milk occurred in late spring and winter periods. Variability in the content of the protein of interest in milk between individuals was noted. During the study a relatively low level of rhLF was established in animal №0747 - 0.11–2.52 g/l, while its maximum value was detected in the producer № 0403 - 4.52–9.06 g/l (more than 4 times difference). The range of individual variability in terms of rhLF content in the milk of goats of the third and the fourth lactation was 0.11–9.06 g/l. Milk of genetically modified goats contains 3 times more lactoferrin than human breast milk. Therefore, rhLF concentration testing in goat milk and tracking the dynamics of this indicator allows us to obtain data that can be used later for breeding pro-grams formation, milk production planning and also to improve the gene pool of transgenic animals.


1994 ◽  
Vol 40 (11) ◽  
pp. 2017-2021 ◽  
Author(s):  
E J Keely ◽  
C Faiman

Abstract We used a time-resolved solid-phase fluoroimmunoassay with a sensitivity of 25 ng/L on 40-fold-concentrated urines to measure urine prolactin (PRL) excretion. The nature of the immunoreactive material was verified to be PRL by: (a) column chromatography showing a monomeric 23-kDa peak; (b) similarity between fluoroimmunoassay and bioassay (Nb2 lymphoma cell) results; and (c) Western blot identification. In 20 normal subjects [serum PRL 6.8 (3.8-14.0) micrograms/L, median (and range)], urine PRL excretion was 0.15 (0.07-0.23) ng/h and 0.24 (0.15-0.54) micrograms/mol of creatinine. Urine values in seven hyperprolactinemic patients were all greater than the upper limit of normal. The correlation between urinary excretion rate and serum values was highly significant (r = 0.979; P &lt; 0.001). These results indicate that a monomeric, immunologically reactive, biologically active form of PRL can be measured in urine at concentrations approximately 0.0005 that in serum. This urine PRL method may provide a practical tool for the repetitive, noninvasive study of PRL dynamics in field studies and in patients with reproductive disorders.


2021 ◽  
Vol 54 (1) ◽  
pp. 34-40
Author(s):  
OM Losev ◽  
HM Voytenko ◽  
NV Kurdil ◽  
MH Povoznikov ◽  
AA Kalashnikov ◽  
...  

Aim. The search for ways to create hepatoprotective food additives based on biologically active substances of plant origin and bee honey is an important area of research in the field of biology and medicine. Materials and Methods. The material for the research was bees of Ukrainian breed (Apis mellifera sossimai). The process of enriching the natural properties of natural honey was carried out in a natural way through the bee family by feeding colonies of bees with a special composition of natural origin components based on Avéna satíva and Silibum marianum. Sieve honey with monofloral honey from white acacia (Robinia pseudoacacia) 55.0–55.0% was used for feeding of the bee families. The first research group was exclusively fed honey water (control). For the second and third experimental groups, Avena Sativa and Milk Thistle Dry Extract were introduced into the main feed, respectively. Preparation of honey water was carried out on distilled water. Each group of bee families was evaluated according to the complex of biological and economic-useful features during the whole study period. The concentrations of active substances in the obtained batches of honey were determined. Ferulic acid was chosen as the active substance for oat, sum of Silibinin A and Silibinin B for milk thistle. The mass concentration of ferulic acid was measured according to European Standards EN 15662:2008 using Gas Chromatography-Tandem Mass Spectrometry following acetonitrile extraction/partitioning and clean-up by dispersive solid phase extraction (Quechers) method. The limits of quantitation of ferulic acid and sum of Silibinin A and Silibinin B were 0.02 mg/kg. The high content (83.3±0.4 mg/kg) of Silibinin flavonoids (A and B), Isosilibin (A and B), Silichristin A, etc. in honey persists. Instead, ferulic acid precipitated in fairly low concentrations of honey (1.13±0.02 mg/kg). Conclusions. The obtained results indicate that the extract of milk thistle is promising for the creation of enriched natural honey with hepatoprotective properties, as the biologically active components of this plant have characteristics that allow to obtain the final product with a high content of active substances. Key Words: honeybee, biological active substances, hepatoprotectors.


1994 ◽  
Vol 72 (04) ◽  
pp. 578-581 ◽  
Author(s):  
T McNally ◽  
S E Cotterell ◽  
I J Mackie ◽  
D A Isenberg ◽  
S J Machin

Summaryβ2 glycoprotein-I (β2GPI), a cofactor for antiphospholipid antibody (aPA) binding, binds to many anionic macromolecules including heparin. The nature of this interaction with heparin is not well understood and its effect on the purported biological functions of β2GPI is unknown.We have examined the interactions of dermatan sulphate (DS) and different pharmaceutical preparations of heparin with β2GPI by crossed immunoelectrophoresis (CIE) and investigated the effect of these agents on plasma levels of p2GPI antigen (β2GPI: Ag) by a standardised enzyme linked immunosorbent assay (ELISA). P2GPI aPA cofactor activity (β2GPI:Cof) was also measured using a modified solid phase an-ti-phosphatidylserine (aPS) ELISA. CIE results confirmed a heparin-β2GPI interaction with unfractionated (UF) heparin. β2GPI:Ag levels were unaffected by any of the preparations investigated. There were no significant differences in β2GPI:Cof activities of the samples containing LMW heparins or DS but levels of β2GPI:Cof were increased in samples containing UF sodium and calcium heparin preparations (0.5 IU/ml Monoparin, p <0.05, and 10 IU/ml Liquemin and Calcipa-rine, p <0.05).


2017 ◽  
Vol 1 (1) ◽  
pp. 23-30
Author(s):  
Fang Rui-ping ◽  
Gao Hong-yan ◽  
Wang Xiao-li ◽  
Yang Xiao-pin ◽  
Li Li

2019 ◽  
Vol 23 (8) ◽  
pp. 860-900 ◽  
Author(s):  
Chander P. Kaushik ◽  
Jyoti Sangwan ◽  
Raj Luxmi ◽  
Krishan Kumar ◽  
Ashima Pahwa

N-Heterocyclic compounds like 1,2,3-triazoles serve as a key scaffolds among organic compounds having diverse applications in the field of drug discovery, bioconjugation, material science, liquid crystals, pharmaceutical chemistry and solid phase organic synthesis. Various drugs containing 1,2,3-triazole ring which are commonly available in market includes Rufinamide, Cefatrizine, Tazobactam etc., Stability to acidic/basic hydrolysis along with significant dipole moment support triazole moiety for appreciable participation in hydrogen bonding and dipole-dipole interactions with biological targets. Huisgen 1,3-dipolar azide-alkyne cycloaddition culminate into a mixture of 1,4 and 1,5- disubstituted 1,2,3-triazoles. In 2001, Sharpless and Meldal came across with a copper(I) catalyzed regioselective synthesis of 1,4-disubstituted 1,2,3-triazoles by cycloaddition between azides and terminal alkynes. This azide-alkyne cycloaddition has been labelled as a one of the important key click reaction. Click synthesis describes chemical reactions that are simple to perform, gives high selectivity, wide in scope, fast reaction rate and high yields. Click reactions are not single specific reaction, but serve as a pathway for construction of simple to complex molecules from a variety of starting materials. In the last few decades, 1,2,3-triazoles attracted attention of researchers all over the world because of their broad spectrum of biological activities. Keeping in view the biological importance of 1,2,3-triazole, in this review we focus on the various synthetic routes for the syntheisis of 1,4-disubstituted 1,2,3-triazoles. This review involves various synthetic protocols which involves copper and non-copper catalysts, different solvents as well as substrates. It will boost synthetic chemists to explore new pathway for the development of newer biologically active 1,2,3-triazoles.


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