In Commemoration of Dr. Mahdokht Pourmansour, an Outstanding Specialist in the Production of the B.C.G. Vaccine in Iran

Background: Post-translational modifications in bioprocessing and storage of recombinant mAbs are the main sources of charge variants. While the profile of these kinds of variants is considered an important quality attribute of therapeutic mAbs, there is controversy about their direct role on safety and efficacy. In this study, the physicochemical and PK properties of separated charge variants belonging to a trastuzumab potential biosimilar were examined. Methods: The APs, BPs, and main variants of trastuzumab were separated and enriched by semi-preparative WCX. A panel of analytical techniques was utilized to characterize the physicochemical properties of these variants. The binding affinity to HER2 and FcγRs and the PK parameters were evaluated for each variant. Results: Based on the results, the charge variants of the proposed biosimilar had no significant influence on the examined efficacy and PK parameters. Conclusion: During the development and production of biosimilar mAbs, studying the effects of their charge variants on efficacy and PK parameters is needed.

In addition to serving as the head of the Microbiology Department at the Pasteur Institute of Iran from 1991 to 1994, Dr. Pourmansour also managed the Microbiology Research Group. Her responsibility as the director of this department was supervising five departments of this research group, including Bacteriology, Parasitology, Medical Entomology, Mycology, Virology, and Rabies ( Figs. 3 and 4).
As a senior research scientist and expert, she established the Department of Biological Product Control at the Pasteur Institute of Iran in 1986. She was a member of the Scientific Publications (1988) and Expert Councils (1987)(1988)(1989)(1990)(1991)(1992) at the same Institute during her service. Dr. Pourmansour was a senior advisor and the first chair of the Scientific Council of the Pasteur Institute of Iran. She has cooperated with the World Health Organization (WHO) as an expert since 1970. She was also responsible for the implementation and policy-making of research in the fight against infectious diseases at the Pasteur Institute of Iran in addition to other universities and the Ministry of Health (1992). In the same year, she also was in charge of setting up the Center for Communicable Disease Control (CDC) at the Pasteur Institute of Iran.
After 38 years of service, Dr. Mahdokht Pourmansour retired on March 26 th , 1994. During the years 1994-2004, she continued to serve in the research planning and contribution to the development of the Pasteur Institute of Iran as a research consultant and member of the scientific council.  who discovered the anti-typhoid vaccine. The Pasteur Institute of Iran succeeded to produce this vaccine in a small laboratory with the minimum facilities. The first B.C.G. vaccine was prepared in October 1947 and then successfully used by the experts of the Pasteur Institute of Iran to vaccinate many children in the Tehran Municipal Nursery. Gradually, by increasing the production capacity at the Pasteur Institute of Iran, it was injected into health centers in Tehran and then other health centers in the country. This vaccine was prepared by Dr. Mehdi Ghodsi and Dr. Mahdokht Pourmansour with care and compliance with all scientific and technical principles. The first batch of B.C.G. vaccine was produced in the form of 2 ml lyophilized vials and placed at the disposal of health and medical centers under the cold -controlled cold temperature, and after being dissolved in the solution made by the Pasteur Institute of Iran, the vaccine was inoculated into the arms of infants. Children were vaccinated twice with this vaccine. The production project of B.C.G. was carried out with the support of the WHO. In 1950, to revise the production of this vaccine, a contract was made with the Pasteur Institute of Paris, and until 1952, vaccination was performed by intracutaneous injection, and since 1953, the vaccine was injected by the intradermal route. The average annual production of B.C.G. vaccine was three million units, and the liquid vaccine was utilized until 1976. Then, with the provision of suitable equipment and place, the lyophilized vaccine was prepared. The stability of a lyophilized vaccine in adverse thermal and environmental conditions is much more than a liquid vaccine [4] .

Production of cholera vaccine in the Pasteur Institute of Iran
In the first fifty years of the establishment of the Pasteur Institute of Iran, at least five cholera epidemics occurred in Iran. Hence, this institute prepared a large number of cholera vaccines. In studies conducted by the teams of the Pasteur Institute of Iran, it was found that the water of some Tehran's aqueducts was contaminated with cholera-causing bacteria. To prevent cholera epidemics in Tehran, the Pasteur Institute of Iran distributed the mineral water from springs around Tehran in closed containers to the public and also disinfected the drinking water of Tehran [4] .
During the cholera outbreaks in Iran and eastern neighboring countries, about 400,000 doses of cholera vaccine were daily prepared at the Pasteur Institute of Iran, and about 24 million doses of cholera vaccine were produced in a limited time. During the cholera outbreak of 1927, due to the high demand of the country for cholera vaccine and insufficient production of the vaccine by the Pasteur Institute of Iran, the part of cholera vaccine required for the country was purchased from Germany. The cholera outbreak occurred again in 1959-60. At this time, The Ministry of Health provided some of the required equipments from Germany to develop vaccine production at the Pasteur Institute of Iran. The Pasteur Institute of Iran managed to prepare about 7-10 thousand doses of the cholera vaccine and then delivered it to the Ministry of Health of Iran. In 1960, during the cholera outbreak in Afghanistan and Pakistan, the Pasteur Institute of Iran produced 5.9 million doses of cholera vaccine for these two countries. In the same year, countries like India, Iraq, Georgia, and Azerbaijan requested to receive the cholera vaccine from Iran. In 1955, when another cholera outbreak swept the country, the Pasteur Institute of Iran and the Razi Institute prepared millions of cholera vaccines. The amount of cholera vaccine production at the Pasteur Institute of Iran in 1970 reached 23 million doses, of which 15 million doses were provided for the Ministry of Health, and 28 million doses were sent to East Pakistan, Turkey, Saudi Arabia, and Ethiopia. Thus, Pasteur Institute of Iran played a central role in controlling cholera epidemics in this region [4] .

Scientific activities
Dr. Pourmansour's specialized fields of activity involved the search of TB and the production of the B.C.G. vaccine. Researchers of the Pasteur Institute of Iran have been preparing B.C.G. liquids according to scientific and technical principles since 1946. Dr. Pourmansour had the responsibility to keep producing this vaccine since 1971, under the supervision of Dr. Mehdi Ghodsi, the former head of the B.C.G. Department who produced the first dry B.C.G. vaccine sample in 1976 [5] . This vaccine produced by the support of WHO, was injected intradermally into all babies in the country. Also, by performing the tuberculin skin test, they studied the emergence of resistance in the vaccinated individuals [6] . Several experts from the Centre for Communicable Diseases at the Ministry of Health and health experts throughout the country, have benefited from her knowledge in the field of TB and the B.C.G. vaccine over the years.
One of the duties of the Pasteur Institute of Iran since its first years of establishment in 1920, was the production of the cholera vaccine [5,6] . The cholera vaccines produced by the Pasteur Institute of Iran were exported to the countries such as Afghanistan, Pakistan, India, Iraq, Turkey, Saudi Arabia, Ethiopia, Georgia, and Azerbaijan, and the institute has played a dominant role in controlling cholera epidemics in these regions. Another activity of Dr. Pourmansour during the managing the Microbiology Department was to supervise the mass production of highquality cholera vaccines [5] .
Dr. Pourmansour also investigated different aspects of salmonellosis. She actively contributed to a comprehensive study attributing to the prevalent salmonellae in Iran over the years 1970 to 1974 [7] . In 1982, she participated in an international project to determine the geographical distribution of Salmonella serotypes, worldwide [8] . At the same time, she also carried out a project to evaluate the relationship between listeriosis and fetal deaths in Iran. One of her latest research was the study of Legionella pneumophila infection of Tehran hospital water samples in 1997 [9] . She also performed several researches on mycoplasma and antiserum production. In 2012, Dr. Mahdokht Pourmansour was honored as one of the top two microbiologists in the country by the Iranian Microbiology Association and received an award from the National Congress of Microbiology held at Tehran.

Scientific trips
In 1957, Dr. Pourmansour spent times at the Statens Serum Institute in Copenhagen, Denmark and the Pasteur Institute of Paris to prepare several samples of the lyophilized B.C.G. vaccine to compare the characteristics of the lyophilized B.C.G. vaccine produced at the Pasteur Institute of Iran with the international standards. She travelled to many countries such as Hungary (Tuberculosis Congress, 1983), the USA (Immunology and Allergy Congress, Chicago, 1992) and France (Listeriosis Congress, 1983, 1985and 1991 to participate in scientific meetings. Dr. Pourmansour had a journey to Alexandria, Egypt and Ethiopia in 1970 to teach the vaccine lyophilization techniques. She went to Syria in 1972 at the invitation of the WHO to set up a laboratory for endemic diseases in this country.

Books and papers
The book "Thirty Years of Continuous Care of the B.C.G. Strain in Iran", published in Persian and French in 1978, is one of Dr. Pourmansour's works [10] . She wrote a chapter in the book "Toxoplasmosis, Tularemia and Listeriosis" in 1983 [11] (Fig. 5) and a chapter "The Application of Tuberculin and B.C.G." from the book "Tuberculosis" in 1987 [12] .

Conclusion
Dr. Mahdokht Pourmansour can be a successful role model for young researchers in Iran. Despite having a medical degree and the chance to continue her clinical work, she chose the field of public health and worked at the Pasteur Institute of Iran to play a significant role in the production of B.C.G. and cholera vaccines to control these diseases in the country. She was strict in overseeing the correct execution of the affairs and narrates that "I tried to send all the employees of the Department of B.C.G. to attend the specialized courses at the Pasteur Institute of Paris or the Copenhagen Institute in Denmark to maintain discipline in working in a standard international production unit". She advises the future-making youth in the country by recommending effort and hope: "We worked hard to build a better future for you; you should do this for posterity. You should be careful not to waste your life's opportunity as far as you can, and you should act in such a way that at the end of your service, you will be satisfied with your performance".