m Animal cell culture is used to

produce variety of commercial &

medicinal prods.
m Evaluation of new drugs on culture
cells is being done which is cost-
effective and less time consuming.
@áhole blood consists of RBCǯs,áBCǯs,platelets
are suspended in fluid called plasma.
@ when blood is allowed to clot it forms cross-
linked network known as serum.
@ Various comp of blood serve a wide range of
physiological func.
@ Specific blood prtns that are of therapeutic
use are:-
clotting factors Serum albumin.
fibrinolytic agents. Ig.
PRODUCT PRODUCED APPLICATION
BY
HGH Mouse mammary Dwrafism,body
tumour cell growth
T-PA Chinese hamster thrombolysis
cell
EPO CHO cells anemia
FSH CHO cells infertility
Factor VIII Baby hamster Hemophilia A
kidney cells
MABs Hybridoma cells Hemophilia B
Factor IX CHO cell lines Hemophilia B
K T-PA is an enzyme produced by
mammalian cells in small amts.
K This removes blood clot and
obstruction in blood vessels.
K First prtn to be produced on a
commercial scale.
K þ
  had cloned human T-PA for
treating blood clots.
K The trade name of this product is
6

Human mRNA coding for T-PA from human
mylenoma cell line.
Reverse transcription
converted to cDNA

Inserted to Chinese Hamster Cell lines

CHO cells excrete T-PA in medium

Isolation of T-PA
4 blood loss must be stopped in case of
injury in which blood vessels are
damaged.ss
4 prtn fibrinogen fibrin.
4 aggregation of platelets at site of injury
stops blood flow
4 blood coagulation process is dependent
on various clotting factors[factor I-XIII].
4 factor VIII prevents hemophilia.
à In hemophiliac patients blood keeps
flowing out in case of injury.
à In 1984,gene that controls production of
F VIII was isolated.
à This enabled research & development of
genetically engineered clotting factor.
à In 1992, 1st genetically engineered
clotting factor was introduced under
trade name R 
6 & R
6
à In 1993, þ
  produced 2nd clotting
factor under trade name
þ
6 .
à There are 4 other recombinant F VIII:-
þ
þ
 R6
 R
R 
 
^EPO is a glycoprtn hormone
produced primarily by cells of
kidney.
^Regulates RBC production.
^Secondary amts of hormone is
synthesized in liver of healthy
adults.
à This sialoglycoprotein hormone consists of 165 amino
acids that form a single polypeptide chain containing
two intra-chain disulfide bonds (Cys7-161 and Cys29-
33) and four potential glycosylation sites, three of
which are
-linked (Asn24, Asn38 and Asn83) and one
-linked (Ser126) (1,2).
à The molecular mass of EPO is 30-34
kDa, but the molecular mass of the
peptide chain is only about 18 kDa. Thus,
about 40% of the glycosylated EPO
molecule consists of carbohydrate,
mostly in the form of
-linked complex-
type glycans.
à Recombinant human EPO (rhEPO)
is produced commercially by the
expression of EPO cDNA clones in
eukaryotic cell lines, most
commonly in Chinese hamster
ovary or baby hamster kidney cells
with CMV and EF-1 promoters
à Transgenic pig containing the the human EPO gene
placed under EF 1 promoter was developed

à F2 generation piglets was analysed for pres of rHu-

EPO
à recovery of rHu- EPO
à two-step precipitation method for the recovery of the
recombinant human erythropoietin (rhEPO·
à ammonium sulfate and divalent metal ions yield was
82.1%
å rHu- EPO is used for variety of clinical
situations
å increased production of RBCs
å anemia in HIV patients.
å curing post operative anemia.
à Albumin from plasma fractionation is generally
considered to be a safe and effective product.
à A number of factors makes recombinant albumin
manufacture increasingly attractive. These factors
are as follows:
m The perceived risk of viral contamination in
serum albumin
m likelihood of reduced availability of recovered
plasma in the future
m price
à Major component of blood plasma
à Fluid balance b/w blood & tissues
à Regulates transport of Amino acids, Fatty acids
and Hormones, Drugs and drug metabolites
^ The expression of human albumin in the milk of
transgenic cows has been identified as the most
promising approach.
à A mammary gland expression vector that
contained the goat Ʌ-casein gene promoter,
5Xupstream regulatory region, exons 1,2, intron 1
as well as the human serum albumin (hALB) mini-
gene (including the full-long sequences of hALB
cDNA and its intron 1).
DIn exhaustible & in expensive.
DFree from infectious agents.
DNew blood prtns not available
earlier .