The V5 epitope tag: technology for vaccines, diagnostics and disease treatment.
Submitting InstitutionUniversity of St Andrews
Unit of AssessmentBiological Sciences
Summary Impact TypeTechnological
Research Subject Area(s)
Medical and Health Sciences: Immunology, Medical Microbiology
Summary of the impact
Proteins are fundamental to life and to many drugs, vaccines and new
types of applied medicine
with engineered cells. For this work, it is often essential to tag
proteins to enable their identification
and purification. The V5 tag, which was developed in St Andrews, is used
very widely in this role
and has some key advantages over alternatives.
Key impacts are:
- V5 tag used in 112 issued patents since 1/1/2008, focussed on
treatment of cancer,
Alzheimer's, viral infection etc.
- The reagents for V5 tag detection had sales exceeding £600k and
generated royalties for
St Andrews of £298k (Jan 2008 to Jul 2013).
- Over 130 different products currently available from commercial
suppliers make use of V5
- Recent vaccine and diagnostics development has relied on V5
In 1993 the group led by Prof. R Randall (in post since 1985), at
the University of St Andrews was
developing methods of new vaccine production targeted at human and simian
viruses (HIV and SIV). This required the presentation of recombinant
proteins to the immune
system to induce an immune response and so they developed a method of
solid matrix-antibody-antigen (SMAA) complexes. These were highly
immunogenic, and were used
to make a vaccine against SIV, to help inform how a vaccine to HIV could
be produced. However,
a method to capture and immobilise the SIV proteins in the SMAA complex
was not available. The
Randall lab had the idea of making recombinant SIV proteins with a peptide
tag attached for
purification and capture purposes to generate SMAA complexes using a
antibody. At the time no such system was commercially available, but the
Randall lab identified a
linear epitope of parainfluenza virus type 5 (originally known as the Pk
tag, later renamed the V5
tag) for which they produced a highly specific monoclonal antibody
(anti-V5). In 1994, they
attached the V5 tag to the p27 protein of SIV and showed that it could be
used with the anti-V5
antibody to construct p27-containing SMAA complexes .
The Randall lab together with the lab of Prof Ron Hay (in post 1985 to
2005) went on to clone the
V5 tag onto many other recombinant proteins. In the period 1995-96, they
general applicability of the V5 tag/antibody combination for the detection
and purification of a wide
range of proteins by a variety of immunological techniques. These included
allows detection of tagged proteins in a cell extract,
immunoprecipitation, which allows the
purification of tagged proteins from cells, and immunofluorescence, which
allows detection of
tagged proteins within cells. Early studies in St Andrews focussed on the
role of protein:protein
interactions in viruses by the Randall lab [2, 3] and in the
ubiquitin signalling pathway by the Hay
lab . Together, this research demonstrated the broad utility of
the V5-tag/antibody across the
spectrum of applications used by both academic and applied scientists
working with proteins.
Subsequent collaborations in 1997-98 further established the utility of
the V5 tag in a wide range of
experimental applications such as yeast cells  and the antibody
was sent to many academic
labs around the world before being commercialised in a deal with Serotec.
In 1999, a series of new
monoclonal antibodies raised against the V5-tag were described, some of
which gave less
background immunofluorescence than the original .
References to the research
St Andrews Authors in bold. Their employment dates were: Bermingham
1993-98; Botting 1993-present;
Dunn 2003-07; Green 1991-95; Hay 1985-2005; Precious 1985-2013; Randall
Ryan 1993-present; Rodriquez 1997-00; Szawlowski 1989-present; Thompson
These are all published in international, peer-reviewed journals. Total
 Hanke, T., Botting, C., Green, A., Szawlowski,
P.W., Rud, E. & Randall, R.E. (1994)
Expression and purification of non-glycosylated SIV proteins, and their
use in the induction and
detection of SIV-specific immune responses. AIDS and Human Retrovirus
Research 10, 653-662.
 Precious, B., Young, D.F., Bermingham, A.,
Fearns, R., Ryan, M. & Randall, R.E. (1995).
Inducible expression of the P, V and NP genes of the paramyxovirus SV5 in
cell-lines and an
examination of NP:P and NP:V interactions. Journal of Virology 69,
 Randall, R.E. & Bermingham, A. (1996).NP:P and
NP:V interactions of the paramyxovirus
simian virus 5 examined using a novel protein:protein capture assay. Virology,
 Roff, M., Thompson, J., Rodriguez, M.S.,
Jacque, J.M., Baleux, F., Arenznan-Seisdedos, F. &
Hay R.T. (1996) Role of I kappa B alpha ubiquitination in
signal-induced activation of NF-kappa B
in vivo. Journal of Biological Chemistry 271, 7844-7850. (179
 Craven, R.A., Griffiths, D.J.F., Sheldrick, K.S., Randall,
R.E., Hagan, I.M. & Carr, A.M. (1998).
Vectors for the expression of tagged proteins in Schizosaccharomyces
pombe. Gene 221, 59-68.
 Dunn, C., O'Dowd, A.M. & Randall, R.E. (1999).
Fine mapping of the binding sites of
monoclonal antibodies raised against the Pk tag. Journal of
Immunological Methods 224, 141-150.
Details of the impact
The V5 tag and antibody system has become an integral part of the
molecular and cellular
biologist's toolkit in industrial, healthcare and commercial laboratories.
From 01/01/2008 to
31/07/2013, the impact of the underpinning research can be seen in
economic terms (>£600k total
sales with £298k in Royalties to St Andrews), commercial R&D (>110
patents issued making use
of the technology) and public health (new vaccines for emerging
multiply-drug resistant bacteria).
Direct economic impact
The V5-tag and anti-V5 antibody were developed for research purposes in
the Randall laboratory
in the mid-1990s. The excellent specificity and utility of this antibody
meant that it was soon in high
demand in the research community. In 1995, a royalty agreement was made
University of St Andrews and the US company, Adb Serotec, under which the
Randall lab would
provide Serotec with the purified anti-V5 antibody in return for 50%
royalties from sales. AbD
serotec also produces a variety of directly conjugated antibodies for
which the University receives
40% royalties. A subsequent deal with Invitrogen (1996) resulted in that
company marketing a
variety of V5 antibodies along with an extensive range of vectors to add
the V5 tag to proteins.
Serotec now purify the antibody directly and supply to Invitrogen under
this agreement. Direct
sales of the V5 antibody and related products covered by the Royalty
agreement amount to £298k
for 2008- 03/2013 [S3]. According to the Senior Vice President of
the company, the V5
monoclonals are "one of (Serotec's) most successful products ... sold
to a large variety of life
science based companies." [S1]
Broader impacts on non-academic R&D
Invitrogen is part of Life Technologies Corporation, a company valued at
$13.6 billion in its
takeover by Thermo Fisher Scientific in April 2013 [S4]. The
importance of the V5 tag/antibody
technology to Invitrogen is evidenced by the fact that the company has
developed over 100
products making use of the technology in their highly successful TOPO and
vectors [S5]. These are marketed and sold to a wide range of
commercial as well as academic
The secondary impacts arising from the use of the V5 antibody in
non-academic research are more
difficult to qualify in absolute terms but there is clear evidence of the
reach and significance of the
V5-antibody in these non-academic spheres:
800 patent applications filed and over 110 patents issued in the period
2008-July 2013 made
explicit use of the V5-epitope for the science underpinning the patent [S6].
Examples from patents
issued in 2012-13 include:
- the inhibition of virus infection (patent US8263570)
- molecular markers for characterization of human
cancer states (patent US8268568)
- engineered organisms with enhanced fermentation
activity to improve chemical product yields (patent
- treatment of cancer with novel monoclonal antibodies
- therapeutic treatments for Alzheimer's disease (patent
The development of vaccination is one of the biggest public health
successes of the past 100
years. New vaccines are required urgently to treat emerging and newly drug
The V5 tag and antibody "have been invaluable tools in the development
of subunit vaccines
against HIV-1 and TB (tuberculosis)" [S2]. A vaccine against
tuberculosis, developed using the V5
tag, is currently undergoing phase IIb clinical efficacy trials in South
Africa. The vaccine, MVA85A,
has already been shown to be "safe and well tolerated" [S7].
This is the first of a new generation of
vaccines against TB and has already completed 15 clinical trials (the
current vaccine, BCG, is not
100% effective and TB causes an estimated 1.4 million deaths a year
according to the WHO).
Diagnostics — advantages of the V5 tag over alternatives
West Nile virus is a serious emerging disease and rapid new diagnostic
methods are needed
urgently. The virus can cause fatal neurological disease, but 80% of
infected humans don't show
any symptoms [S8]. Therefore it is important to develop methods to
detect the virus in blood
samples. In 2011 a new diagnostic reagent based on the prM antigen was
reported, which is
suitable for the detection of antibodies against West Nile virus in serum
samples. This requires
expression of the prM antigen in tagged form so that it can be
immobilised. Use of a polyhistidine
tag "disrupted" the protein so that it was not useful as a
diagnostic antigen. Use of the V5-tag
however "allowed formation of the authentic antigenic structure and the
proper presentation of the
V5 epitope", allowing the development of a "useful diagnostic
agent" [S9]. Thus, although a variety
of tags are available, the V5 tag has clear advantages in some
Sources to corroborate the impact
[S1] Email from the Senior vice president, Serotec. Corroborates
success of V5 products.
[S2] Email from an independent scientific expert, The Jenner
Institute, University of Oxford.
Corroborates utility of V5 technology in vaccine design.
[S3] Audited financial statement showing royalties accruing to St
Andrews University for V5
antibody sales in period 2008- July 2013 of £298,367.
[S4] Widely reported, e.g. http://www.cnbc.com/id/100641197.
Corroborates value of the Invitrogen
[S5] Invitrogen products with V5 tag. Invitrogen website reports
105 products with V5 tag (correct
as of 29 Oct 2013).
[S6] Patent search using Google, corroborates number of patent
applications and awards using
this technology in the REF period.
Search patents issued or filed in period 2008-July 2013 with search term
[S7] Details of MVA85A vaccine incorporating V5 tag. Corroborates
use of V5 tag in TB vaccine
under late stage clinical trial.
[S8] Corroborates importance of development of new diagnostics for
West Nile virus.
[S9] Expression of recombinant West Nile virus prM protein fused
to an affinity tag for use as a
diagnostic antigen. Setoh Y.X. et al., (2011), Journal of Virological
Methods 175, 20-27.
Corroborates the claim that the V5 tag has key advantages over other tags
in some circumstances.