QUESTIONS FOR PAPER “A”
Question 1: Research problem
The problem addressed by the research is the scorpion envenoming which is a primary challenge in the most part of the universe. Additionally, the provision of an adequate remedy by health services to this problem has not been easy because the treatment of scorpionism is too complicated. Therefore, the research is addressing the possible treatment to this disease by considering various experiments carried out by using polyclonal antiserum that has been disinfected from the serum of hyperimmune horse (Muyldermans et al 179).
Question 2: Adjuvant utilization
In this research, Freund’s adjuvant was utilized. This was done by mixing unpolished venom H. lepturus with an equivalent volume of Freund’s adjuvant. This process used this mixture plus two 1-yr-old Camillus dromedaries to perform the entire immunization. This adjuvant was used because it is less procedural and also it is affordable in general (Muyldermans et al 179).
Question 3a: The immunization schedule
In this research, the vaccination of camel is designated with raw venom extracted from H. leptus. It has been proved that a desirable proportion of heavy-chain antibodies from polyclonal have the ability to neutralize the scorpion toxic effects. This involved the choosing of HNc-specific as well as Nbs through displaying the phage as well as their effects on the neutralization process. This process was then followed by ELISA process, and the analysis was carried out by using Western blot. This chosen immunization is much involving since the procedures to be followed are numerous, hence high affinity of B-cell may not be generated.
Question 3b: using Material and Method Approach
The methods used under materials is the most procedural method necessary for this research (Muyldermans et al 179). The Nbs were measured through a competitive assey by basing it on the ELSA technique. This was done by coating ninety-six microplates with two distinct HNc concentrations throughout the night at a temperature of 4˚C. This measurement method was useful in that accurate results were recorded at the end of the experiment. By using Beatty formula in calculating the Nbs affinity, precise end results were found.
Question 4: Definition of LD100
According to Compendium of Chemical Terminology (1997), LD100 is the absolute lethal dose. Which is the smallest amount of chemical or else biological substances which is likely to cause a death of the animals being tested on the conditions defined by the technicians. It does vary in connection with animal type as well as administration route. LD50 is a median lethal dose. It is commonly used to kill only half of the tested animals. Hence, a control test can also be performed (Muyldermans et al 179).
Question 5: A diagram of ELISA procedure
Question 6: A diagram of Western Blot
The lymphocytes were made from 100 ml anti-coagulated blood of the immunized camels by utilizing Lymphoprep as indicated in protocol of the manufacturer. The VHH library was built by isolating RHA from the lymphocytes’ pool of the camels by the help of RNA extraction kit. This was followed by converting RNA to cDNA by the help of reverse transcriptase with the oligo primer (Yardehnavi et al 4002). Finally, PCR nesting was carried out to amplify the gene fragments of VHH.
Question 7b: In vivo neutralization of assays
This was done by determining the minimum necrotizing dose of HNc via the use of intradermal injection of raising amount of toxins into the three rabbit back derma. During the injection, digital snapshots were taken and measured at time intervals. The capability of HNc-specific Nbs to neutralize the action of dermonecrotic was evaluated by in vivo method (Yardehnavi et al 4002).
QUESTIONS FOR PAPER “B”
Question 1: Gene rearrangement of camels
An antigen binding portion of camalid HCAbs comprises of one domain referred to as variable domain of HCAbs. It has been shown that HCAbs from one humped camels which have been infected with trypanosomes have the capability to associate tightly as well as specifically to parasite antigens. Thus roughly, equal HCAbs levels as well as H2L2 antibodies are available in the blood of camel. A phylogenetic analysis have shown that HCAbs dedicated Y-gene are sourced from Y-genes coding froe traditional antibodies (Yardehnavi et al 4002). Therefore, HCAbs have been articulated after a V-D-J rearrangement and has to be dedicated to constant gamma-genes. This raises an immune response of camel.
Question 2: Definition of VHH and Nbs
VHH is the main antigen binding stem of HCAbs and is available at in a single amino acid stretch while Nbs are Nano-bodies which have given a greater value as therapeutic molecules as well as tools for clinical diagnostic. The advantages of Nbs includes; they are the smallest ever known antibodies fragments and they have the capability of binding with very high specificity. Nbs possess properties that makes them more superior than other antibodies (Yardehnavi et al 4002). Additionally, they have better tissues penetration properties and they are manufactured at a lower cost. Some of their applications comprises; enzyme inhibition due to their binding capabilities, in therapeutic to treat and diagnose cancer and also to treat tumors due to high tissue penetration properties.
Work cited
Muyldermans, S., et al. "Camelid immunoglobulin’s and nanobody technology." Veterinary immunology and immunopathology 128.1 (2009): 178-183.
Yardehnavi, Najmeh, et al. "A camelid antibody candidate for development of a therapeutic agent against Hemiscorpius lepturus envenomation." The FASEB Journal 28.9 (2014): 4004-4014.
