Selection of downstream steps by analysis of protein surface property: A case study for recombinant human lactoferrin purification from milk of transgenic cow

Zhang, Yan; Luo, Jian; Zhang, Songping; Bai, Qian; Li, Zenglan; Li, Qiang; Liu, Yongdong; Zhang, Guifeng; Ma, Guanghui; Su, Zhiguo

doi:10.1016/j.procbio.2015.05.025

Cited by 7 publications

(3 citation statements)

References 37 publications

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“…HPLC is one of the key techniques for a simple and low-cost protein characterization [ 14 , 16 ]. Size-exclusion HPLC (SEC-HPLC) is mainly used for the evaluation of Lf aggregation, integrity, and purity [ 10 , 17 , 18 ]. Reversed-phase HPLC (RP-HPLC) methods are suitable for the qualitative and quantitative analysis, of various Lf samples.…”

Section: Introductionmentioning

confidence: 99%

Stability-Indicating Analytical Approach for Stability Evaluation of Lactoferrin

et al. 2021

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Lactoferrin is a multifunctional iron-binding glycoprotein in milk. Due to its potential for the treatment of various diseases, interest in products containing lactoferrin is increasing. However, as a protein, it is prone to degradation, which critically affects the quality of products. Therefore, the main purpose of our work was to develop a stability-indicating analytical approach for stability evaluation of lactoferrin. We were focused on two complementary methods: reversed-phase and size-exclusion chromatography. The stability-indicating nature of the selected methods was confirmed. They were successfully validated by following the ICH guidelines and applied to preliminary lactoferrin stability studies. Up to three degradation products, as well as aggregates and fragments of lactoferrin, were detected in various samples using complementary reversed-phase and size-exclusion chromatographic methods. The analytical approach was additionally extended with three spectroscopic techniques (absorbance, intrinsic fluorescence, and bicinchoninic acid method), which may provide valuable complementary information in some cases. The presented analytical approach allows the stability evaluation of lactoferrin in various samples, including the ability to detect differences in its degradation mechanisms. Furthermore, it has the potential to be used for the quality control of products containing lactoferrin.

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Section: Introductionmentioning

confidence: 99%

Stability-Indicating Analytical Approach for Stability Evaluation of Lactoferrin

et al. 2021

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“…Furthermore, the preparation of antigen and antibody is complicated and expensive, and it is not suitable for the testing of large quantities of samples. Enzyme-linked immunosorbent assay (ELISA) is more selective and sensitive; the ELISA kit has been used for bovine lactoferrin determination [20,21,22]. The commercially available ELISA kit is not suitable for camel lactoferrin determination, however, because the target protein cannot bind the antigen and antibody in the kit.…”

Section: Introductionmentioning

confidence: 99%

Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard

Xu³

et al. 2019

Molecules

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An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of lactoferrin in camel milk based on the signature peptide. The camel lactoferrin was purified by heparin affinity chromatography and then used to screen tryptic signature peptides. The signature peptide was selected on the basis of sequence database search and identified from the tryptic hydrolysates of purified camel lactoferrin by ultrahigh-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry. The pretreatment procedures included the addition of isotope-labeled winged peptide and the disposal of lipids and caseins followed by an enzymatic digestion with trypsin. Analytes were separated on an Acquity UPLC BEH 300 C18 column and then detected on a triple-quadrupole mass spectrometer in 7 min. The limits of detection and quantification were 3.8 mg kg−1 and 11 mg kg−1, respectively. The recoveries ranged from 74.5% to 103.6%, with relative standard deviations below 7.7%. The validated method was applied to determine the lactoferrin in ten samples collected from Xinjiang Province.

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“…Take the GRASP2 program for example, it is a graphics program to view macromolecules, which is particularly written for the display and manipulation of the surfaces of molecules and their electrostatic properties [15]. Zhang et al reported the protein surface analysis with the GRASP2 program helpful for rational comparison and selection of chromatographic resins for efficient design of purification schemes [16,17], demonstrating the possibility and efficacy of rational design based on understanding of fundamentals. Although interaction-predictive models lead to exceptional levels of process understanding, their requirements for well-known characteristics of all the proteins limit their use for developing separations of complex and unclear mixtures.…”

Section: Introductionmentioning

confidence: 99%

Selection of downstream steps by analysis of protein surface property: A case study for recombinant human lactoferrin purification from milk of transgenic cow

Cited by 7 publications

References 37 publications

Stability-Indicating Analytical Approach for Stability Evaluation of Lactoferrin

Stability-Indicating Analytical Approach for Stability Evaluation of Lactoferrin

Determination of Lactoferrin in Camel Milk by Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Using an Isotope-Labeled Winged Peptide as Internal Standard

Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis

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