Native Protein Separation by Isoelectric Focusing and Blue Gel Electrophoresis-Coupled Two-Dimensional Microfluidic Chip Electrophoresis

Hou, Ni; Yu, Chunshui; Yu, Shiyong; Quan, Zongliang; Pan, Chenhua; Deng, Yulin; Li, Geng

doi:10.1007/s10337-014-2728-3

Cited by 7 publications

(3 citation statements)

References 18 publications

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“…First, a free solution based two-dimensional chip electrophoresis for native protein separation was established based on our previous work. 21 A mixture of carbonic anhydrase, catalase and myohemoglobin were chosen to constitute a protein mixture for electrophoresis separation. The simultaneous analysis of myohemoglobin and creatine kinase is regarded as reliable and valuable for the early diagnosis of acute myocardial infarction (AMI).…”

Section: Development Of the Ief/cze Coupled Two-dimensional Chip Elec...mentioning

confidence: 99%

A novel method to predict protein aggregations using two-dimensional native protein microfluidic chip electrophoresis

Xu²,

Huang

et al. 2016

Anal. Methods

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Section: Development Of the Ief/cze Coupled Two-dimensional Chip Elec...mentioning

confidence: 99%

A novel method to predict protein aggregations using two-dimensional native protein microfluidic chip electrophoresis

Xu²,

Huang

et al. 2016

Anal. Methods

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“…Assuming the premise that protection from A. marginale transmission was due to conformational protein epitopes is correct, Western blotting after native (i.e., non-denaturing) PAGE is another possibility for identification of protective protein antigens. Limitations of native PAGE include (1) proteins are not separated based solely on size and (2) less information is available about the use of native gels for the second dimension of 2D-PAGE Arndt et al, 2012;Lasserre and Ménard, 2012;Hou et al, 2014). Furthermore, it is not certain that 2D-PAGE would not render protective antigens non-reactive with immune sera.…”

Section: Discussionmentioning

confidence: 99%

Interference with biologic transmission of Anaplasma marginale by targeting the tick-pathogen interface

Shahzad¹

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[EMBARGOED UNTIL 6/1/2023] Ticks are globally renowned vectors for numerous zoonoses. Ixodid ticks transmit a variety of viral, bacterial, and protozoan pathogens to their vertebrate hosts. Because of the expansion of the geographic ranges of both ticks and pathogens, increasing tick populations, emerging tick-borne pathogens and continued challenges of achieving effective and sustained tick control, ticks and tick-borne diseases have increased in importance from medical and veterinary public health perspectives. Host immune resistance to ticks has been studied since the first half of the 20th century. This dissertation addresses (i) adaptation of a naturally coevolved large-animal model to investigate immunological underpinnings of resistance to tick challenge, (ii) interference with experimental biologic transmission of a naturally coevolved tick-borne pathogen, Anaplasma marginale, to cattle, (iii) preliminary observations of effects of tetracycline on acquisition and transmission of A. marginale by tick vectors and (iv) preliminary optimization of artificial feeding conditions for ixodid ticks with different apparatuses. Several tick proteins were identified, which were reactive to antisera of tick-extract-immune cattle that were associated with reduced tick performance. The proteins identified here are important for future experiments to determine their posited roles in reduction of tick feeding. Dermacentor andersoni and D. variabilis ticks, natural biologic vectors of A. marginale in the United States, did not experimentally transmit this pathogen to susceptible hosts immunized with crude homogenates of tick tissues. In addition to the main emphasis of this dissertation, preliminary observations included reductions in infection levels acquired by ticks and effects of tetracycline on tick acquisition and transmission of A. marginale. Collectively, the work described in this dissertation suggested that interventions with acquisition and transmission of A. marginale and other tick-borne pathogens are not necessarily dependent on tick mortality.

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“…In Blue Native PAGE, a negative charge is added to proteins by the addition of Coomassie dye allowing for separation of proteins in a native state. A limitation with these approaches will be the two-dimensional separation of these proteins as there are limited protocols that describe two-dimensional separation of proteins while conserving native structure (350)(351)(352). This work could potentially involve significant effort to discover and optimize a protocol for native two-dimensional gel electrophoresis.…”

Section: Native Protein Identificationmentioning

confidence: 99%

Refinement of an established large-animal model to understand the tick-pathogen-host interface

Hoffman¹

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Ticks are globally distributed vectors of important pathogens of human and animal health. Since the discovery of tick resistance in 1918, the field has sought continuously for the development of effective biologic control of ticks and tick-borne pathogens. A goal of this dissertation was the refinement of a large-animal model system to study species indigenous to the United States. Further, the research described in this dissertation sought to target various aspects of the tick-pathogen-host interface. The research in chapter 2 detailed our work in high-quality sequencing of the non-tick-transmissible Anaplasma marginale Illinois strain. We identified several candidate genes and genomic elements associated with the tick-transmission of A. marginale. The research in chapter 3 sought to adapt and refine a large-animal model system to evaluate host resistance to Dermacentor andersoni ticks. Several proteins isolated from cattle with reduced tick performance were identified in calves immunized with tick salivary gland tissues. The proteins identified here are important for future experiments to determine their posited roles in reduction of tick feeding. Finally, the research in chapter 4 built on this work and suggested that immunization with native salivary gland (NSG) and native midgut (NMG) interfered with A. marginale transmission by D. andersoni. Further, NSG immunization also had the most consistent and greatest impact on D. andersoni acquisition of A. marginale. The antigens responsible for this protection remain undetermined. The results in this dissertation demonstrate that there are several facets of the tick-pathogen-host life cycle that can be targeted for mitigation of tick-borne disease.

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Native Protein Separation by Isoelectric Focusing and Blue Gel Electrophoresis-Coupled Two-Dimensional Microfluidic Chip Electrophoresis

Cited by 7 publications

References 18 publications

A novel method to predict protein aggregations using two-dimensional native protein microfluidic chip electrophoresis

A novel method to predict protein aggregations using two-dimensional native protein microfluidic chip electrophoresis

Interference with biologic transmission of Anaplasma marginale by targeting the tick-pathogen interface

Refinement of an established large-animal model to understand the tick-pathogen-host interface

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