Human Cell Lines for

Biopharmaceutical
Manufacturing

History, status, and future

perspectives
Presenters

 Samra Mustafa
 Amina Azam
 Attiya Kamaal
Points to be discussed
01 INTRODUCTION
An overview

02 HISTORY
 Non-human expression systems used to manufacture
biotherapeutic products
 Human cell lines used to manufacture licensed products
03 APPLICATIONS
 Clinical and pre-clinical development by HCL in
the expression of proteins

04 RISKS Vs BENEFITS
Future perspectives
Conclusion
 Introduction
HUMAN
CELL LINE
Human cell lines
are immortalized
cells propagated in
vitro from primary Biopharmaceutical
explants of human
tissue or body fluid. Manufacturing
Pharmaceutical manufacturing is the
process of industrial-scale synthesis of
pharmaceutical drugs as part of the
pharmaceutical industry.
Series of unit operations:
 Milling
 Granulation
 Coating
 Tablet pressing
 Protein therapeutics (including
PROTEIN monoclonal antibodies, peptides and
recombinant proteins) represent the

THERAPEUTIC largest group of new products in

development by the
biopharmaceutical industry.

Mammalian Expression
System
It is the system of choice for studying the
function of a particular protein in the most
physiologically relevant environment.
Non-mammalian Expression
System
Expression systems are genetic constructs
that are designed to produce a protein, or
an RNA outside a cell other than
mammalian cells.
Comparison
RECOGNITION EXPRESSION PROCESSING SIGNALS

Recently developed Biotherapeutic proteins

Toady most biotherapeutic proteins developed to date have been
produced using the mammalian expression system as in
Chinese hamster ovary and murine myeloma (NS0, Sp2/0) cell
Mammalian expression system

lines, and now there has been a recent shift toward the use of
human cell lines.
No
n-M

Expression Accumulation
am
ma

System is having difficulty Accumulation of proteins

lian

in expressing some as inclusion bodies

mammalian proteins.
Ex
pre

Contamination Endotoxin
s
sio

Protease contamination This system has a

n

from host proteins leading disadvantage of

to degradation of endotoxin accumulation.
expressed protein
What Among The non-human mammalian cell lines
produce PTMs that are not expressed in

Mammalian
humans, namely galactose-α1,3-
galactose (α-gal) and N-
glycolylneuraminic acid (NGNA)

Expression System Non-human cell lines are usually

screened during their production to
identify clones with acceptable glycan
profiles
 CHO
HUMAN CELL
01 LINES
 BHK21
 NSO & Sp2/0

Human cell lines produce proteins like

NON-HUMAN
the humans.
In particular, the structure, number and CELL LINE 02
location of post-translational N-glycans
can affect the biologic activity, protein
stability, clearance rate and
immunogenicity of biotherapeutic
proteins
 HeLa
 HT 1080
 PER.C6
 HISTORY

2014 2021
1960s 2012
1951

HeLa, was Human diploid Taliglucerase alfa rFVIIIFc and Directed

established in cells were was rFIXFc are differentiation of
1951 developed in the approved by the recombinant human induced
from a cervical 1960s for vaccine US FDA for fusion proteins pluripotent stem
cancer manufacturing the treatment of that cells to hepatic
type 1 Gaucher were approved by stellate cells
disease. the FDA
 NHES Used To Manufacture
Biotherapeutic Products
Many non-human expression systems have been utilized in
the production of currently approved biotherapeutic 0
proteins

Expression systems and their role in the manufacturing of

biotherapeutic products

The most frequently used mammalian system is the CHO

cell line, which is used in the manufacture of >70% of
currently approved recombinant proteins

For proteins produced in all these non-human cell lines, as

well as those produced in human cell lines, potential safety
concerns arise from the possibility of process-related
contaminants and immunogenicity
Human cell lines used to manufacture
licensed products
HEK293
Human cell line most often
used in the production of
biotherapeutic proteins,
which offer the advantage
HT-1080
of producing fully human HT-1080, was produced from a
PTMs fibrosarcoma with an epithelial-
like phenotype With the use of
gene activation technology four
approved therapeutic proteins
have been produced
by Shire

PER.C6
This system was originally
developed to produce
human adenovirus vectors
for use in vaccine
development and gene
therapy
 Overall Comparison
Human Cell Line Other Expression

80% 20%

ADVANTAGES DISADVANTAGES

 Clinical experience is not as extensive as for

o Absence of potentially immunogenic PTMs due
other cell lines, although experience is
to human-compatible glycosylation
growing
o Easily grown in suspension serum-free culture
 Potential susceptibility to human viral
o Achieve rapid reproduction
contamination
o Amenable to several transfection methods
 Clinical
APPLICATIONS Or
Pre-Clinical
MOR103
MOR103 is in clinical development for the
treatment of rheumatoid arthritis and
multiple sclerosis.

CL184
In a phase 1 clinical study, it demonstrated a favorable
safety profile and rapid development of rabies virus
neutralizing activity

PER.C6
Currently, a variety of products utilizing the PER.C6 cell
line are in phase 1 or 2 clinical trials

Clinical Development
 HKB-11

Two additional Cell lines are

utilized by products currently in
preclinical development.

“
Pre-Clinical
Development
Human CAP cell lines
 Risks Vs Benefits
Observational
Studies

Number of patients and their health sta-

Option A Option B Option C Option D
tus ration
Number of patients and their health sta-
Option A Option B Option C Option D
tus ration

All Clinical Trials Interventional Studies

70% 80%
 BENEFITS RISKS
 Human cell lines can
serve as a valuable niche
expression system for  A theoretical concern
biotherapeutic proteins with the use of
 Achieving equal human cell lines is
productivity to other
mammalian cell lines
 The production of
Vs an increased risk of
transfer of human
adventitious agents
proteins that lack
potentially immunogenic
 In the future, with additional research
investments and a continuation of the
technologic advances that have
already led to improvements in the use
of human cell lines for
protein manufacture, human cell lines
FUTURE
will be further optimized
More sophisticated product collection PERCPETIVES
strategies
will be developed, and these cell lines
may become one of the
preferred platforms for protein
biotherapeutic production.
CONCLUSION

The recognition of these prospects

and issues should be regarded as
quite a healthy set of actual
opportunities for the treatment of
untreatable disease as well as
production of useful therapeutic
proteins as this the need of our
time.
REFERENCES

 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5404623/
 https://www.nature.com/articles/s41596-021-00509-1
 https://www.nature.com/articles/s41536-020-00100-4
 https://www.atcc.org/products/crl-12568
 https://www.cell.com/trends/biotechnology/fulltext/S0167-
7799(20)30244-4
 https://spectrum.ieee.org/the-human-os/biomedical/
devices/qa-ai-could-redesign-the-drug-development-
process
THANK YOU