VINTAGE PAPER

Introduction
Fermentation and cell culture are at the heart of drugs and
biologics manufacturing. However, unlike most manufac-
turing processes in contemporary electronics or automobile
industries, manufacturing in the biotechnology industry is
not well-dened. Indeed, it has been observed that processes
for manufacturing potato chips are more robust than those
for proteins. In part, the complexity of living systems is
responsible for this situation. The other part is that tools for
process monitoring are not as widely available in order to get
a better understanding of the complex relationship between
gene expression and environmental factors. In order to obtain
consistent production from bioreactors, it is necessary to
carefully monitor and control the process. Consistent scale-
up also requires that bioreactor operation conditions be kept
uniform across different scales. One of the most common
scale-up parameters employed is the volumetric oxygen
transfer coefcient, or k
L
a which essentially is a measure
of how much oxygen can be supplied to cells growing in a
bioreactor. Since oxygen is typically the limiting nutrient
due to its poor aqueous solubility, in general cells grown in
two dissimilar bioreactors (e.g., a shake ask and a stirred
tank) but operated at equal k
L
a will show similar growth and
product formation kinetics (Gupta and Rao, 2003).
However, until the advent of dissolved oxygen electrodes,
measuring k
L
a was not easy. In contrast to prior methods,
the Bandyophadhyay article provided a simple and elegant
means to dynamically measure k
L
a (Bandyophadhyay et al.,
1967). Only a fast responding dissolved oxygen probe
was needed and the technique used was based on a brief
interruption in the air (oxygen) supply to the bioreactor
and recording the subsequent dissolved oxygen trace. The
technique was rapidly adopted and became textbook
material (Pirt, 1975). Several articles followed over the
years to improve on the original method. Fred Heineken
published a article that carefully considered the quantitative
effect of probe response time on the accuracy of the k
L
a
measurement (Heineken, 1971). A few years later, Hill and
Robinson (1974) showed how the simple technique could be
further extended to estimate specic growth rate of the
culture. A further study by Tribe et al. (1995) led to
quantitation of the errors resulting in k
L
a measurements if
probe response time was accounted for. This entire body of
work is now being applied to the quantitative determination
of k
L
a in present day bioreactor systems such as bag systems
and minibioreactors that use non-invasive oxygen sensors
(Hanson et al., 2009).
Here is what Professor Arthur Humphrey himself had to
say via email about his perspective on the article. He recalls
that Professor Taguchi and I wanted to devise a way to
measure oxygen transfer rates in small and medium sized
fermenters in order to calculate k
L
as and then to be able to
correlate these coefcients with agitation and aeration rates
plus fermenter design. This was in the days before on-line
computers. He then goes on to say that My motivation for
the concept was that you only needed a fast response oxygen
probe (the MIT group had developed a cheap and easy way
for students to make these probes) to make the measure-
ments. Hence, any laboratory could make oxygen transfer
rate measurements and these could be compared with
growth rate measurements. In my subsequent visits to
various biotech labs throughout the worldJapan, China,
India, Taiwan, Australia, etc., I was amazed at how many
laboratories were using the technique. After publication at
least two dozen articles appeared rening the theory and
dening its application limits. One of the best follow-up
articles was written by Fred Heineken.
So there it isthis article, by virtue of its impact on the
practice of modern biochemical engineering, is a true classic
and is one that continues to stand the test of time as the
technology for bioreactors and oxygen measurement evolves.
Govind Rao
Center for Advanced Sensor Technology &Department of
Chemical and Biochemical Engineering
University of Maryland, Baltimore County
Baltimore, Maryland
References
Bandyopadhyay B, Humphrey AE, Taguchi H. 1967. Dynamic measure-
ment of the volumetric oxygen transfer coefcient in fermentation
systems. Biotechnol Bioeng 9:533544.
Gupta A, Rao G. 2003. A study of oxygen transfer in shake asks using a
non-invasive oxygen sensor. Biotechnol Bioeng 84:351358.
Hanson M, Brorson K, Moreira AR, Rao G. 2009. Comparisons of optically
monitored small-scale stirred tank vessels to optically controlled dis-
posable bag bioreactors. Microbial Cell Factories 8:44.
Heineken FG. 1971. Oxygen mass transfer and oxygen respiration rate measure-
ments utilizing fast response oxygen electrodes. Biotechnol Bioeng 13:599618.
Hill GA, Robinson CW. 1974. Measurement of aerobic batch culture
maximum specic Growth rate and respiration coefcient using a
dissolved oxygen probe. Biotechnol Bioeng 16:531538.
Pirt SJ. 1975. Principles of microbe and cell cultivation. London: Blackwell
Scientic.
Tribe LA, Briens CL, Margaritis A. 1995. Determination of the volumetric
mass transfer coefcient (k
L
a) using the dynamic gas out-gas in
method: Analysis of errors caused by dissolved oxygen probes. Bio-
technol Bioeng 46:388392.
Correspondence to: Govind Rao, Center for Advanced Sensor Technology & Depart-
ment of Chemical and Biochemical Engineering, TRC, UMBC, Baltimore, Maryland
21250; telephone: 410-455-3415; fax: 410-455-1049; e-mail: grao@umbc.edu
Published online in Wiley InterScience (www.interscience.wiley.com).
DOI 10.1002/bit.22566
2009 Wiley Periodicals, Inc. Biotechnology and Bioengineering, Vol. 104, No. 5, December 1, 2009 841
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