immunization-alpacas-lama-pacos-with-protein-antigens-production

Immunization of Alpacas (<em>Lama pacos</em>) with Protein Antigens and Production of Antigen-specific Single Domain Antibodies

In this manuscript, a method for the immunization of alpaca and the use of molecular biology methods to produce antigen-specific single domain antibodies ...

University of Kentucky

We recently found that brain tissue from patients with type-2 diabetes (T2D) and cognitive impairment contains deposits of amylin, an amyloidogenic hormone synthesized and co-secreted with insulin by pancreatic β-cells. Amylin deposition is promoted by chronic hypersecretion of amylin (hyperamylinemia), which is common in humans with obesity or pre-diabetic insulin resistance. Human amylin oligomerizes quickly when oversecreted, which is toxic, induces inflammation in pancreatic islets and contributes to the development of T2D. Here, we tested the hypothesis that accumulation of oligomerized amylin affects brain function.

Neuroinflammation and neurologic deficits in diabetes linked to brain accumulation of amylin.

 This study describes the normal variations in serum and cervicovaginal fluid (CVF) cytokine levels throughout pregnancy.

Patterns of Systemic and Cervicovaginal Fluid Inflammatory Cytokines throughout Pregnancy.

Glucan phosphatases are a unique subset of the phosphatase family that bind to and dephosphorylate carbohydrate substrates. Family members are found in diverse organisms ranging from single-cell red algae to humans. The nature of their functional oligomerization has been a source of considerable debate. We demonstrate that the human laforin protein behaves aberrantly when subjected to Size Exclusion Chromotography (SEC) analysis due to interaction with the carbohydrate-based matrix. This interaction complicates the analysis of laforin human disease mutations. Herein, we show that SEC with Multi-Angle static Light Scattering (SEC-MALS) provides a method to robustly define the oligomerization state of laforin and laforin variants. We further analyzed glucan phosphatases from photosynthetic organisms to define if this interaction was characteristic of all glucan phosphatases. Starch EXcess-four (SEX4) from green plants was found to lack significant interaction with the matrix and instead exists as a monomer. Conversely, Cm-laforin, from red algae, exists as a monomer in solution while still exhibiting significant interaction with the matrix. These data demonstrate a range of oligomerization behaviors of members of the glucan phosphatase family, and establish SEC-MALS as a robust methodology to quantify and compare oligomerization states between different proteins and protein variants in this family.

Article	Year
Neuroinflammation and neurologic deficits in diabetes linked to brain accumulation of amylin. Molecular neurodegeneration\| PubMed ID: 25149184	2014
Patterns of Systemic and Cervicovaginal Fluid Inflammatory Cytokines throughout Pregnancy. American journal of perinatology\| PubMed ID: 29132178	2018
Oligomerization and carbohydrate binding of glucan phosphatases. Analytical biochemistry\| PubMed ID: 30291838	2018

Savita Sharma

Biography

JoVE Journal Publications

Other Publications