Reference no: EM132751321
Background infoIxodes scapularisI. scapularisBorrelia burgdorferi
Climate change means the range for the , also known as deer ticks or black legged ticks is expanding. The reason why this is important is because carries the etiologic agent of Lyme disease, .
Lyme disease was first identified as a juvenile arthritis in Lyme, CT. At first, it was seen as an autoimmune disorder, as some arthritis cases are, but later the B. burgdorferi the spirochete that caused of the disease was isolated.
A vaccine was developed in 1998, but pulled from the market after 3 years.
You now work for Dr. Gary Deeney, renowned expert on B. burgdorferi and interested in patenting a new vaccine. You are in the early stages of preparing a grant and a presentation to Baroness Research Ltd, with the goal of developing a new Lyme disease vaccines in humans, and in cattle.
B. burgdorferi is a Gram negative Spirochete, that has a very reduced genome that suits it's parasitic lifestyle. It cannot synthesize any of the amino acids, and lack other genes considered to be part of primary metabolism. It doesn't have TCA or an electron transport chain, it does have a phosphotransferase system (group translocation) to import glucose. It doesn't have many traditional virulence factors, noted virulence factors include the outer membrane surface proteins, two of noted importance include OspA and OspC. Designated as lipoproteins, these ARE NOT the same thing as animal lipoproteins. These proteins are imbedded in the outer membrane of the organism, though often associated with one face (inside or outside) of the outer membrane. Other virulence factors, such as CspA, CspZ and OspC inhibit complementation.
QUESTION
The potential funders of this project, Baroness Research Ltd. want Dr. Deeney to demonstrate the vaccine generate an adaptive immune response in cattle after initial testing is complete.
Design an ELISA, or something close to an ELISA, that can assay for this vaccine's efficacy.
For this hypothetical proposal, you have unlimited supplies of
- ELISA plates
- ELISA reagents
- purified CspZ,
- Escherichia coli that can express CspZ
- anti-CspZ antibodies that were raised in mice
- Enzyme-linked Rabbit anti-mouse antibodies
- whole blood and serum from cattle exposed to Borellia burgdorferi
- whole blood and serum from cattle not exposed to B. burgdorferi
- whole blood and serum from cattle exposed to your vaccine candidate then B. burgdorferi
- whole blood and serum from cattle not exposed to Borellia burgdorferi, but given your vaccine
- Enzyme linked Chicken anti-cattle antibodies
- Anti IFNgamma antibodies
- Enzyme linked anti IFNgamma antibodies
When thinking about this, Baroness Research wants to to demonstrate that a humoral response to CspZ is found in response to your vaccine.
This is a digram, not an essay, so draw out this assay stepwise. Give a clear demonstration to Baroness Research that you know what your are talking about and can communicate that to them.