- Preparation and evaluation of in house monoclonal antibodies against commercial antibodies for clinical and research use
- Intravenous immunoglobulin (IVIg) preparation
- Human Leukocyte Antigen (HLA) Typing
- In house production of diagnostic kit (direct agglutination test(DAT)) for the visceral leishmaniasis diagnosis
- In house production of trans-isolate media(TIM) for the transport of cerebrospinal fluids for primary cultures from patients with bacterial meningitis
- In house production of viral load and drug resistance testing for different viral diseases such as HIV, viral Hepatitis etc
- Developing flow cytometry based diagnostics for leukemia
- Genomics and Bioinformatics Research
Brief summary of some of the current projects
Preparation and evaluation of in house monoclonal antibodies against commercial antibodies for clinical and research use
Monoclonal antibodies are antibodies designed in the laboratory to recognize and bind to specific proteins. These antibodies have immense use in research, diagnostics and therapeutics. Currently, a third of new drugs that are introduced make use of monoclonal antibodies.
In this project, we are involved in locally producing laboratory scale mouse anti-human CD4 and CD38 monoclonal antibodies (mAb) using hybridoma technology. Anti-CD4 monoclonal antibodies can replace commercialized antibodies to monitor treatment response to anti-retroviral therapy of HIV patients. CD38 is a prognostic marker for lymphocytic leukemia. Therefore, anti-CD38 monoclonal antibodies can be used to monitor the prognosis of lymphocytic leukemia. The project is meant to transfer the technology of mAb preparationand fluorescence labeling by using local expertise and international partners. Three complementary approaches are used for this project i.e., in house preparation of hybridomas, production of monoclonal antibodies from purchased hybridomas, and labeling purchased monoclonal antibodies.
Intravenous immunoglobulin (IVIg) preparation
Intravenous immunoglobulin (IVIg) is sterile IgG prepared by purifying human plasma pooled from large number of donors. IVIG is composed of more than 95% unmodified IgG, which has intact Fc-dependent effector functions, and trace amounts of immunoglobulin A (IgA) or immunoglobulin M (IgM). IVIg is mainly used to treat people with immunodeficiency disorders and sometimes it is co-administered with other medications for people who are fighting chronic infections. The demand for IVIg is increasing globally. Personal communication with medical doctors in Ethiopia has revealed that IVIG is also needed in this country; however, scientific studies assessing the need for IVIg in Ethiopia have not been conducted yet.
The IVIg purification project was initiated with seed money from AHRI. Following that, ministry of health of Ethiopia donated a significant amount of money from their sustainable development goals (SDG) fund to facilitate IVIg technology transfer and laboratory scale production.
Human Leukocyte Antigen (HLA) Typing
Kidney transplant is the best long-term therapeutic strategy for individuals with end stage kidney disease. In some transplant cases, there could be humoral or cellular mediated immune responses against the transplanted kidney, even in the presence of immunosuppressants. This leads to organ rejection, which leaves the patient in a worse medical condition. Most rejection reactions are directed at human leukocyte antigens (HLAs) expressed on the transplanted kidney. In order to avoid rejection during kidney transplant, multiple HLA compatibility tests must be done for the patient-donor pair.
This project will provide compatibility tests for the kidney transplant program at St. Paul’s Hospital. The three types of compatibility tests we do are serology typing, molecular typing, and flow cytometry crossmatch. Until now, these tests were done at a company in Germany or India and it took two weeks to get the results, which was unpromising for patients that were at the end stages of kidney disease; so is the need for local capacity of doing the tests in Ethiopia. AHRI took the initiative and trained its staff at the University of Michigan and established HLA laboratory at AHRI. In collaboration with St Paul Kidney transplant program, validation has been done with few samples tested simultaneously at both our facility and at the company in Germany. All our results were concordant with the results from the German company.
In house production of trans-isolate media (TIM) for the transport of cerebrospinal fluids for primary cultures from patients with bacterial meningitis
The Bacterial and Viral Diseases Research Directorate is currently conducting research to determine prevalence and serogroup distribution of meningococcal meningitis among vaccinated and non-vaccinated Ethiopians. As a part of this study, it has been initiated to produce trans-isolate media (TIM), which supports survival of meningitis strains during transport to the laboratory. Most bacterial causes of meningitis remain unconfirmed because of failure to grow isolates in culture due to the distance from laboratories. Bacterial causes of meningitis die easily outside of the host and require enriched media and body temperature to grow. TIM provides nutrients for live transportation. The technology for in house TIM production has been acquired so that quality assured readymade vials of TIM is provided for clinicians and researchers.
Genomics and Bioinformatics Research
A team of scientists from AHRI have secured a grant for a project called TBGEN, which will study host-parasite interactions in tuberculosis. We are using this project to establish and build genomics and bioinformatics research at AHRI. In order to prepare for the project, we have already started giving training to staff and students at AHRI. One of the trainings we gave on bioinformatics is described briefly below.
Bioinformatics training at AHRI
Institutional initiatives strengthening capacity at AHRI focuses on building a bioinformatics training center, next generation sequencing (NGS) facility and computing platform to support researchers and postgraduates in Ethiopia. The facilities benefit both ongoing and new project initiatives such as on pathogen evolution, virulence determinants and epidemiology of important pathogens, including M. tuberculosis.
Training workshop on Mycobacterium whole genome sequence data analysis
As part of capacity building in bioinformatics in Ethiopia, an introductory course on bioinformatics was conducted between 24- 28 December 2018 at AHRI. The training was aimed at delivering hands on practical introduction to NGS data analysis of Mycobacterium tuberculosis complex (MTBC) genome. Every workshop day included, 40 minutes presentation, three hours hands on practical and 20 minutes discussion. The presentation topics were on next generation sequencing technologies, examples of sequence data file formats and a stepwise description of each bioinformatics workflow during NGS data analysis.
There were 12 participants (7 from AHRI, 1 each from EPHI, NHADIC, AAU-IBT, EBTI, and AASSTU) and 4 trainers (2 from AHRI, one from AAU-IBT and one from EBTI) from health and biotechnology research institutes in Ethiopia.
At the end of the training, participants had an opportunity to reflect on the training experience. Though they have expressed their satisfaction on acquiring basic skills in handling NGS data, they have highlighted the need to improve computational power of the workstations at the bioinformatics laboratory and address fluctuations of internet connectivity.