An Ultra Bad Hair Day – Stratagene
May 1, 1998
N

Ecco Hair Design
999 Encinal Lane
San Diego, CA 92037
Dear Ecco Hair Design,
I am writing to express my disgust with your new
shampoo, “UltraHair.” My modeling career (which
includes high profile shoots in magazines such as
the Active Lifestyles swimwear edition and Glam
Magazine) has been nearly ruined by your product.
After using your shampoo, I suffered severe split
ends and a flaky, sore scalp. It took several
weeks of treatment under the supervision of a
dermatologist to return my hair to normal (although
it still lacks the luster it once had). Your
claims that this product would provide deep
conditioning benefits are appalling.
As a result of this experience, I had to cancel
several shoots, losing over $100,000 in income and
potentially losing additional shoots in the future.
Because of this grave inconvenience and direct
negative impact on my career, I have contacted my
attorneys to take legal action against your
company.
You will hear from my attorneys soon. In the
meantime, I urge you to remove this product from
the market at once.
I would appreciate your immediate attention to this
matter.
Sincerely,
Natalie
Natalie Georgette
P.S. My attorneys advised that I should provide you
with the batch number for the offending shampoo:
0298/12.
Willy, “UltraHair” product coordinator at Ecco Hair Design, stared in disbelief at the letter he
held in his hands. The production manager, Maria, had given it to him as he arrived in his office
after his lunch break. Although Maria was ultimately responsible for the production of the
shampoo, she believed that Willy was the best person to determine what had gone wrong. Willy
coordinated the activities of the team involved in all stages of “UltraHair” production.
This incident was serious, unlike cases with other products in which a customer had been mildly
dissatisfied and merely requested a refund. Willy wondered, Did the problems stem from a
mistake in the initial research and development stages? Or did something go awry when the
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An Ultra Bad Hair Day – Stratagene
shampoo was produced on a large scale? Why hadn’t the quality control team identified the
problem? Were there other issues to consider? Willy and his team would need to identify the
cause of the problem, why it was not discovered before the shampoo was released for sale, and
how the problem should be fixed. Was the problem isolated to a specific batch, or must Ecco do
a nationwide recall? This was potentially a high-profile case, since it involved a well-known
model. The threat of a lawsuit made solving the problem Willy’s highest priority.
What potential causes should Willy consider?
Based on the symptoms described by Natalie, Willy ruled out the possibility of incorrect
detergent concentration (this particular concentration had been used successfully with other
similar products). Moreover, the scents involved in production had been thoroughly tested by
another company prior to their inclusion in the shampoo.
Willy began by calling a meeting of the team within Ecco Hair Design who had been involved in
the making of “UltraHair.”
The Structure of Ecco Hair Design
The “UltraHair” team included individuals from Research, Process Development,
Manufacturing, and Quality Control (see Appendix for a flowchart of potential departments
within biotechnology companies). The responsibilities of these divisions are as follows.
The Research Division is where the Ecco Hair Design scientists developed the initial idea for
“UltraHair.” Mark works in this division. After completing a Master’s Degree in molecular
biology, he was fortunate to find a job in a company where he could explore his own ideas and
turn them into an actual product. He and his research group determined the ingredients needed to
make “UltraHair” shampoo extra conditioning.
In Process Development, Ann was responsible for scaling up the research process used in
making “UltraHair” from the laboratory bench level to a pilot plant level. This included scaling
up the fermentation process used to prepare protein additives by working with the fermentation
division—a division whose staff members often received an overwhelming number of orders at
one time. Ann’s Bachelor’s Degree in molecular biology provided her with the skills necessary
for these responsibilities.
Within the Manufacturing Division, Steve supervised the group that produced the final product.
In doing this, he followed stringent procedures (Good Manufacturing Practice, or, GMP) to
ensure a consistent product. This required a background in science and the mechanical skills to
work with sophisticated equipment. Fortunately, Steve was certain that he had completed the
necessary paperwork and documentation during the actual manufacturing of “UltraHair.” This
would allow him and his colleagues to review all steps he had taken during manufacturing.
In the Quality Control (QC) Division, Sharon ensured that “UltraHair” met certain minimum
specifications of quality such as purity of ingredients. To confirm this purity, she used an
approved QC protocol. Sharon received her training through a biotechnology program at a local
community college.
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An Ultra Bad Hair Day – Stratagene
Put yourself in Willy’s shoes... If you were Willy, what questions would you
ask Mark, Ann, Steve and Sharon, based on the role that she or he had in the
making of “UltraHair”?
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An Ultra Bad Hair Day – Stratagene
Willy’s Next Steps
Together with Mark, Ann, Steve and Sharon, Willy reviewed the chemicals and general steps
taken in developing the shampoo. The shampoo contained the following ingredients:
Ingredient
Purpose
Purified Water
Solvent
Sodium Laurel
Sulfate
Detergent
Protein
Luster, manageability,
conditioning
Coloring &
Fragrance
Aesthetics
Methylcellulose
Thickening agent
Upon seeing that the shampoo contained
proteins, Willy asked his team whether protein
contaminants could cause Natalie’s symptoms.
The team decided to investigate whether a
contaminant had found its way into the desired
protein additive. Yet the process Ecco used to
separate proteins had been fool-proof to date.
Did the separation process work? Did an
undesirable ingredient remain in the shampoo?
If so, how?
The Process of Protein Purification
First, what is a protein? Proteins are chain-like molecules
composed of amino acids. Every protein has a unique
structure and 3-dimensional shape and unique chemical
properties. In the case of “UltraHair,” protein additives
were intended to make hair silky and strong while
providing conditioning benefits for the scalp. (See
Appendix and Resource List for additional
information about proteins.)
How do you isolate a protein and where do proteins come
from? Protein purification is essentially a method of
separating the desired species of protein out of a mixture
of proteins. The separation is done based on the proteins’
physical or chemical properties such as size or charge.
One method of purifying a protein is to pass the protein
through a column packed with a resin whose chemical or
physical properties allow each protein in a mixture to
interact with it in a different manner. This method is
referred to as “column chromatography.” By passing the
proteins through a column, some of the proteins will pass
through, and some will be retained on the resin. There are
hundreds of resins, each one with unique properties. By
choosing the appropriate resins, a specific protein can be
separated from all the others in a mixture.
The protein is collected in a vial as it passes through the
column. This collected amount of protein is referred to as a
“fraction.” (See Appendix for more details regarding
protein separation.)
At first glance, it seemed that the proper steps had been taken to make the proteins for the
shampoo. Mark outlined the purification process that he had developed in research, sharing his
results which showed that a contaminant protein was, in fact, removed by the process.
Ann reported the steps she had taken to scale the process up. She had used almost exactly the
same protocol that Mark had developed, with only minor modifications. In particular, Ann had
retained the same molecular sizing resin (G25) that Mark had used in the final purification step,
which is the important step for removing the contaminant protein. G25 resin excludes all
molecules whose size is greater than 5,000 daltons molecular weight (MW).
Steve produced the actual batch records from the manufacturing runs, proving that all of the steps
had been performed to specifications.
Finally, Sharon affirmed that all the quality control assays had been done correctly, including the
final spectrophotometric analysis of the G25 column purification step (see Figure 1). Figure 1
shows the spectrophotometric analysis of the fractions collected off of a G25 sizing column.
Fractions 9–27 (the left peak which appears red in color in the collected fraction) had been
collected because they contained the desirable protein additive for the shampoo (the “luster
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An Ultra Bad Hair Day – Stratagene
protein”); fractions 42–63 (the right peak which appears yellow in color in the collected fraction)
had been discarded because they contained a protein contaminant (a protein that was known to
cause reactions such as scalp irritation).
1.4
1.2
ABS 450 nm
1
0.8
0.6
0.4
0.2
75
66
57
48
39
30
21
12
3
0
Fraction #
Figure 1. Spectrophotometric analysis of the G25 column purification step.
[insert photograph of column]
As Willy and his team reviewed the documents, they discovered that a new batch of protein
additive had been prepared by fermentation and used in the production lot of shampoo used by
Natalie Georgette. The batch preparation was purified according to Standard Operating
Procedures (SOPs). SOPs are the step-by-step guidelines that a company develops to ensure that
the exact same steps are taken each time a product is reproduced. The SOP also serves as a
record of the steps taken in making a product so that in cases like this, the steps can be reviewed
to detect where a problem may have arisen. The final G25 column separated a possible
contaminant from the desired protein additive. According to the quality control assays, which
used the red color of the protein additive as a measure of its relative purity, the materials
obtained at this step were clean for use in production. Documentation was complete. See Figure 2
below for a graphic representation of the protein separation process used in making the original
formulation for “UltraHair.”
Protein mixture
containing both
desirable and
undesirable
proteins
Known
Contaminant
Discarded
Separated using
G25 column
“Luster
Protein”
Used in
production of
“UltraHair”
Figure 2. The Protein Separation Process Used in the Original Formulation of “UltraHair.”
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An Ultra Bad Hair Day – Stratagene
What Now?
The team’s findings appeared to put him at another dead end, but Willy persisted. He decided to
check the purity of the new batch in a different way, by using another assay method called SDSPAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Willy thought, “Fortunately,
we freeze small samples from the fermentation run.” He loaded the SDS-PAGE gel with a known
set of molecular weight standards (Lane A) and the two separate aliquots from the fermentation
runs (Fermentation 0298/121 in Lane B and Fermentation 0697/082 in Lane C). After staining the
gel, he anxiously looked at the results (see Figure 3).
Figure 3. SDS-PAGE results. [NEED TO REPLACE]
This could be it! The two batches were not identical. In addition to the desired product, an extra
protein band was visible in the new batch preparation.
Now Willy must clean up (remove the unwanted band) the batch of protein additive. His own
research work had not yielded a contaminant in the material recovered after the G25 sizing step,
yet a contaminant was clearly visible on the SDS-PAGE gel.
Willy has decided to try to separate the two proteins by using a different size exclusion resin
(G50; this excludes all molecules whose size is above 30,000 dalton molecular weight, MW) and
an ion exchange resin (which separates proteins on the basis of differences in charge) with SDSPAGE analysis to visualize all proteins. He tells you—his research associate—that this is your
task.
What do you need to know to carry out Willy’s instructions? This is your
chance to ask questions to clarify what Willy and his team discovered.
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2
“Fermentation 0298/12” refers to the batch that was fermented in February (02), 1998 (98) in Fermenter #12 (/12).
“Fermentation 0697/08” refers to the batch that was fermented in June (06), 1997 (97) in Fermenter #8 (/08).
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