1 / 11

The Purification and Characterization of bFGF-like Human Autoantibodies

The Purification and Characterization of bFGF-like Human Autoantibodies. By Veena Venkatachalam. What is a neuron?. Neurons – cells that make up our nervous systems have processes that transmit electrical impulses (action potentials)

toyah
Download Presentation

The Purification and Characterization of bFGF-like Human Autoantibodies

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. The Purification and Characterization of bFGF-like Human Autoantibodies By Veena Venkatachalam

  2. What is a neuron? • Neurons – cells that make up our nervous systems • have processes that transmit electrical impulses (action potentials) • this is how your brain communicates with the rest of your body • neurodegenerative diseases cause these neurons to die • basic fibroblast growth factor (bFGF) promotes neuron survival

  3. What is bFGF? • bFGF – growth factor that stimulates elongation and branching of neuronal processes • Why don’t we use bFGF to treat patients with Alzheimer’s or Parkinson’s disease? It degrades too rapidly! • Solution – find a stable substance that has bFGF-like effects

  4. What is an antibody? • Antibodies – Y-shaped proteins that are produced by cells of the immune system in response to a stimulus; protect us by neutralizing toxins • four chains – two heavy chains and two light chains • each chain has one variable region; variable regions differentiate antibodies from one other and determine specificity

  5. Complementary Our Hypothesis • We have hypothesized, based on past observations, that serum from several patients contains an antibody that has effects similar to those of bFGF Complementary bFGF (neurotrophic growth factor) Idiotype (antibody complementary to bFGF) Anti-idiotype (bFGF-like antibody; similar antigen binding site)

  6. Hydroxy-apatite gel Immuno-affinity column Isolating the Antibody: Purification Methods • We used three different methods to purify the patient serum: Protein A beads: bind IgGs Immunoaffinity column: has anti-bFGF antibodies bound to the protein A beads; thus, bFGF like antibodies bind very strongly. Elute w/ diff. pH Hydroxyapatite (HA) column: pour protein A eluate over column, elute w/ diff. molarities of sodium phosphate Protein A beads: pour serum over column and elute with citric acid; result => only IgGs

  7. Bioactivity Assays • Bioactivity assays were used to determine the fraction that we wanted to purify further

  8. Acknowledgements I’d like to extend my deepest appreciation to Dr. Zimering for his guidance. I would also like to thank Robert Donnelly for his help in running mass spectroscopy and amino acid sequencing our samples.

More Related