he DIVISION of BIOLOSICS STANDARDS Jiflj-stbtnljj Congress of % ®nitrtr Stotts of 
the fptst jfressura, 
Began and held at the City at Washington on Monday, the second day of December, one 
thousand nine hundred and one. 
AJN" ACT 
To regulate the sale of viruses, serums, toxins, and analogous products in the 
District of Columbia, to regulate interstate traffic in said articles, and for 
other purposes. 
Be it enacted by the Senate and House of Representatives of the United 
States of America in Congress assembled, That from and after six months after 
the promulgation of the regulations authorized by section four of this Act no 
person shall sell, barter, or exchange, or offer for sale, barter, or exchange in 
the District of Columbia, or send, cany, or bring for sale, barter, or exchange from 
any State, Territory, or the Distrust of Columbia into any State, Territory, or the 
District of Columbia, or from any foreign country into the United States, or 
from the United States into any foreign country, any virus, therapeutic serum, 
toxin, antitoxin, or analogous product applicable to the prevention and cure of 
diseases of man, unless (a) such virus, serum, toxin, antitoxin, or product has 
been propagated and prepared at an establishment holding an unsuspended and 
unrevoked licence, issued by the Secretary of the Treasury as hereinafter authorized, 
to propagate and prepare such virus, serum, toxin, antitoxin, or product for sale in 
the District of Columbia, or for sending, bringing, or carrying from place to place 
aforesaid; nor (b) unless each package of such virus, serum, toxin, antitoxin, or 
product is plainly marked with the proper name of the article contained therein, 
the name, address, and license number of the manufacturer, and the date beyond 
which the contents can not be expected beyond reasonable doubt to yield their 
specific resn**- n-*>mded, That the suspension ' - ’’ 
mat~ with the pro>«*ona o» 
this Act be, and the asms are hsrsoy, ispaaltl 
Speaker Bourn qf Representatives. 
fJ * -A llt g.M m jyuk AMUMM 
4riWwlw» Wf OMW the DIVISION of BIOLOGICS STANDARDS 
“To apply our present knowledge to the im- 
provement of biologies now on the market, 
to find better ways of producing and testing 
these biologies, and to help in the develop- 
ment of new immunizing agents against the 
infectious diseases that, so far, have baf- 
fled science-these are the ultimate aims 
of those concerned with the control of 
biologies.” 
Director, 
Division of Biologies Standards 
For sale by the Superintendent of Documents, U.S. Government Printing Office 
Washington, D.C. 20402 - Price 15 cents DBS staff works in two modern, interconnected 
buildings on the campus of the National Institutes of 
Health, at Bethesda, Maryland. The staff of approximately 
300 includes more than 100 professional members- 
physicians, biochemists, virologists, biologists, and im- 
munologists. Although the research interests of the Di- 
vision cut across many disciplines, the work is organized 
into seven laboratories: Control Activities, Bacterial Prod- 
ucts, Biophysics and Biochemistry, Blood and Blood 
Products, Pathology, Viral Immunology, and Virology and 
Rickettsiology. BIOLOGICS REGULATION 
The DBS is responsible for establishing and maintain- 
ing standards of quality and safety of all biological prod- 
ucts that come within the jurisdiction of the Public Health 
Service. These products include all vaccines, antitoxins, 
therapeutic serums, allergenic products, and human 
blood for transfusion, as well as products prepared from 
human blood. 
Because many of these products are derived from liv- 
ing organisms, such as bacteria and viruses, and all by 
their nature are either potentially dangerous or ineffec- 
tive if improperly prepared and tested, close surveillance 
of production is essential. The development of realistic 
standards for these products and the exercise of proper 
control over them is effectively backed by a flexible re- 
search program. 
The standards for the production and testing of bio- 
logical products are set forth in regulations prescribed 
by DBS. As scientific knowledge advances, revisions are 
made in the regulations. 
Following determination by the DBS that the pre- 
scribed standards for safety, purity, and potency have 
been met, licenses for the manufacturing establishment 
and for the product are issued by the Department of 
Health, Education, and Welfare upon recommendation of 
the Surgeon General of the Public Health Service. 
Food and Drug Administration regulations require the 
monitoring of clinical trials of all investigational drugs. 
The Division therefore maintains surveillance of all ex- 
perimental biological materials under study for potential 
commercial use. CONTROL TESTING 
The safety, purity, and potency of all biologies must be 
established before licensing. Control tests range from 
relatively simple sterility tests to complex and costly 
potency determinations and safety tests. Once a product 
is licensed, continuing surveillance is maintained by 
inspection, review of manufacturing records of produc- 
tion and testing, and by testing samples of the product. 
Animals play an important role in control testing. Mice, 
guinea pigs, rabbits, and monkeys provide a variety of 
responses by which the safety, purity, and potency of 
these products can be evaluated. 
Safety tests in animals are performed to ensure the 
relative freedom from harmful effects to the recipient of 
the product. The guinea pig is used in safety tests for all 
vaccines, toxins, and toxoids. 
Closely allied to safety is the requirement of purity, or 
freedom from extraneous matter in the finished product. 
For example, to ensure freedom from fever-producing 
substances of bacterial origin for antitoxins, plasma, and 
other blood derivatives, a test dose of the product is in- 
jected into the ear veins of rabbits. Elevation of body 
temperature indicates the presence of impurities. 
To determine potency-the specific ability of a bio- 
logic to effect a specific result-a number of specialized 
tests are involved, including animal tests. In diphtheria 
and tetanus toxoid potency tests, the animal may serve 
to measure immune response to the product. The po- 
tency of typhoid vaccine is measured by immunizing 
mice; potency of smallpox vaccine is tested by the rabbit 
skin test. 
Approximately 35,000 control tests are conducted an- 
nually in DBS laboratories on samples of manufacturers’ 
products to ensure their safety, purity, and potency. If 
the DBS test results confirm those of the manufacturer, 
that particular batch of the product can be released for 
commercial use. The DBS tests serve as a basis for the 
annual release of some 4,000 production lots. 
6 Some 180,000 mice, 3,500 rabbits, 2,000 monkeys, and 
8,600 guinea pigs are used annually for control tests. The availability and use of viral and bacterial antigens 
has for many years protected millions of people against 
infectious diseases. Smallpox vaccination and the multi- 
ple antigen (diphtheria and tetanus toxoids and pertussis 
vaccine) are routinely given in this country to children 
of pre-school age. Travelers in areas of the world where 
typhoid fever, yellow fever, cholera, and typhus are preva- 
lent are immunized against these diseases. 
Despite the long-time use of these immunizing agents, 
there is always need for their improvement in the light of 
current scientific knowledge. DBS investigators continue 
to devise better methods of testing for viral, bacterial, and 
rickettsial antigens. 
It was, however, the advent of tissue-culture-produced 
virus vaccines which revolutionized biologies production. 
The first of these was the inactivated Salk polio vaccine, 
licensed in 1955. The development of the live, oral, Sabin 
poliovirus vaccine presented DBS with a new technical 
challenge. The monkey neurovirulence tests required for 
this vaccine involved extensive studies. More than 150 
neurovirulence tests were made in more than 7,500 mon- 
keys during 1959-1961. These studies led to the estab- 
lishment of a standardized test system which assured 
the safety of the Sabin polio vaccine. Research, testing, 
and the development of standards in connection with 
measles vaccine represented another major concern for 
DBS, beginning soon after measles (rubeola) virus was 
isolated by Dr. John Enders in 1954 and continuing until 
the vaccine was licensed in 1963. 
Research is also directed toward the formulation of 
standards for new products. Advances in virology have 
led rapidly toward the development of viral vaccines 
for mumps, German measles, and some of the respiratory 
diseases. Studies are also conducted on diseases for 
which etiologic agents have not yet been isolated. Of 
increasing importance is the intensive study of the role 
that viruses may play in human malignancies. 
VIRAL . BACTERIAL RESEARCH TUMOR VIRUSES 
Many if not all animals harbor latent viruses, some of 
which may have a tumor-producing or “oncogenic” po- 
tential. This is of concern in biologies control since most 
virus vaccines are produced in animals or chick embryo 
tissue cultures or in embryonated eggs. 
One such virus, SV-40, found to be present in monkey 
kidney tissue culture, can cause tumors in hamsters. 
Another, avian leukosis virus, causes tumors in chickens. 
Since the role of these viruses in relation to human 
disease is unknown, the only position the Public Health 
Service can take is to require their exclusion from all 
licensed vaccines. 
Future vaccines will undoubtedly be produced in 
other tissue culture systems; therefore, DBS scientists 
are developing methods and special techniques to reveal 
potentially oncogenic viruses. These include animal in- 
oculation, electron microscopy, and histologic evaluation 
of tumor cells. For example, newborn hamsters have been 
found to be particularly susceptible to tumor induction 
by animal viruses and are used in demonstrating the 
tumor-producing capability of certain of these viruses. 
These studies are important for the entire field of 
cancer research, but are of special concern for vaccine 
safety. When an oncogenic virus is a contaminant of the 
vaccine, it can be eliminated by changing production 
techniques, but if the vaccine virus itself has an on- 
cogenic potential, the problem is infinitely more complex. 
It takes many months of study involving large numbers 
of animals to determine the possible effects of such 
viruses. 
10 DBS uses over 25,000 hamsters annually 
in its research and testing programs for 
oncogenicity. 
Electron microscopy is also a valuable aid 
in searching for and identifying extraneous 
virus particles in cell cultures used in the 
production of vaccines. BIOLOGICS STANDARDIZATION 
Since most biological products cannot be standard- 
ized by chemical or physical means, their strength or 
potency must be tested in relation to established stand- 
ards, or reference preparations. 
The official physical standards for biological products 
in the United States, provided by law and regulations, are 
those established by the National Institutes of Health. To 
ensure that biological products made by several manu- 
facturers will have uniform potency, the DBS provides 
manufacturers with these official standards. Their de- 
velopment depends on advanced knowledge in the fields 
of immunology, biology, virology, biochemistry, and medi- 
cine. More than 7,000 of the preparations prepared and 
tested are sent annually to manufacturers and others 
engaged in biologies standardization. 
DBS scientists are constantly working to improve 
these preparations by developing more accurate and 
economical laboratory tests. For example, complex bio- 
logical products are subjected to electrophoretic frac- 
tionation in order to identify the active components 
which will serve as more specific standards. DBS scien- 
tists are experimenting with stabilizers and freeze-drying 
techniques to develop better methods of preserving and 
storing biological products so that potency will be main- 
tained. Studies are being carried out to establish stand- 
ards of potency for new products pending licensing and 
for substances for which no suitable potency tests have 
been developed, such as allergenic products, poison ivy 
extracts, and certain bacterial vaccines. 
There are compelling reasons why biological stand- 
ards should be uniform throughout the world. This is best 
accomplished through international cooperation. DBS 
scientists serve on World Health Organization expert 
committees involved in the establishment of interna- 
tional standards, and assist other countries in establish- 
ing their own national standard preparations. BLOOD RESEARCH AND CONTROL Research in this field is designed to strengthen con- 
sol procedures for the manufacture and storage of blood 
and blood derivatives. 
During the past ten years, there has been a great in- 
crease in the use of such blood products as normal 
serum albumin, immune serum globulin, normal human 
Plasma, frozen single-donor plasma for coagulation de- 
fects, and sera for determining blood groups and types, 
as well as whole blood for transfusions. 
DBS scientists study the effects of freezing, thawing, 
ar>d drying of these various elements of blood; the effect 
°f storage conditions-temperature, time, light, and con- 
feiner materials-on the properties of whole plasma and 
fee separate plasma proteins. 
Of the estimated 5,000 blood banks in this country, 
230 are operating under Federal license to ship human 
blood from one state to another for commercial use. 
These blood banks account for about 65 percent of the 
approximately 6,000,000 pints of blood collected an- 
nually. The remainder is collected by local blood banks 
for use within the community, and the medical proce- 
dures used are subject to local and state laws and 
regulations. 
Blood banks licensed under the biological products 
control provisions of the Public Health Service Act must 
comply with standards set by DBS. To assure continued 
compliance with these standards, every licensed blood 
bank is inspected annually by DBS personnel, and tests 
are performed on random samples of blood selected by 
the inspectors. INSPECTION 
To ensure continued compliance with Public Health 
Service regulations, specially trained members of the 
DBS scientific staff annually inspect every establishment 
licensed to manufacture biologies, as well as those ap- 
plying for license. 
* U. S. GOVERNMENT PRINTING OFFICE : 1968 O - 284-418 There are more than 200 establishment licenses and over 1,300 product licenses in effect, covering 
approximately 290 specific products. These include: 
Antitoxins 
Immune serum globulins 
Blood and blood derivatives 
Bacterial vaccines 
Toxoids 
Toxins 
Viral and rickettsial vaccines 
Diagnostic substances for skin tests 
Allergenic extracts 
Antivenins 
Poison ivy extracts 
Venoms 
photos: Roy Perry and 
Jerry Hecht THE NATIONAL INSTITUTES OF HEALTH 
PUBLIC HEALTH SERVICE 
U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE 
Public Health Service Publication No. 1 744