RBMOnline - Vol 5. No 3. 259–264 Reproductive BioMedicine Online; www.rbmonline.

com/Article/643 on web 23 August 2002

Articles
New developments in gonadotrophin
pharmacology
Dr Gordon is currently Director of Reproductive Medicine at Organon Pharmaceuticals Inc,
in West Orange, New Jersey. He earned his doctorate from Monash University, Australia,
and did his postdoctoral work at the Jones Institute in Norfolk, Virginia. Dr Gordon has
received many awards and honours in his areas of expertise and is actively involved in
several professional societies. Dr Gordon has published over 60 original journal articles,
reviews, book chapters, and book reviews in his field. He has also presented his work at
many national and international meetings.

Dr K Gordon

Keith Gordon, Associate Director, Reproductive Medicine, Organon Pharmaceuticals Inc, 56 Livingston Avenue – 4th
Floor, Roseland, NJ 07068, USA
Correspondence: e-mail: k.gordon@organoninc.com

Abstract
This article reviews the past, present, and future of gonadotrophin therapy, including purification of gonadotrophins from
animal or human urine sources and production of gonadotrophins through recombinant technology. With the advent of
recombinant DNA methodologies combined with site-directed mutagenesis, a variety of structural modifications becomes
possible.

Keywords: corifollitropin alfa, follicle-stimulating hormone, FSH-CTP, gonadotrophins, human chorionic gonadotrophin,
luteinizing hormone

Introduction could transmit diseases such as Creutzfeldt–Jakob disease

The word gonadotrophin is derived partly from the Greek
word tropé? (turning on) and is used to describe hormonal
agents that initiate and sustain gonadal functions. In humans,
there are three gonadotrophins: follicle stimulating hormone
(FSH), luteinizing hormone (LH), and chorionic
gonadotrophin (CG). The gonadotrophins (FSH, LH, and CG)
together with thyroid-stimulating hormone (TSH) are
glycoproteins, so named because there are a number of
carbohydrate (sugar) residues attached to certain amino acids
in their protein backbones. The gonadotrophins and TSH are
all heterodimers, with each hormone consisting of a common
α-subunit and a hormone-specific β-subunit (Pierce and
Parsons, 1981).
Systematic investigations into the existence of gonadotrophins
began in the 1920s, when the surgical technique of
hypophysectomy was developed and studies in animals
quickly led to the realization that removal of the pituitary
gland led to gonadal regression and loss of reproductive
capacity (Smith, 1926). Reproductive function could be
restored by administering suitably prepared water-soluble
extracts of the pituitary gland (Fevold et al., 1931). The first
clinical use of exogenous gonadotrophin to induce ovulation in
women was achieved in the 1950s with pituitary extracts
(Gemzell et al., 1958). However, the use of this cadaveric
source material was stopped because it became apparent that it
(Lunenfeld, 2002).
Fortunately, gonadotrophins extracted from pituitary glands
were soon superseded by human menopausal gonadotrophin
(HMG) products extracted from the urine of post-menopausal
women (Lunenfeld, 2002). However, the LH activity of the
raw material (urine of post-menopausal women) is only
approximately one-third of that of the FSH activity in terms of
IU. Accordingly, it is necessary to supplement the LH activity
with human CG (HCG) derived from the urine of pregnant
women in order to meet the pharmacopoeial requirements for
HMG preparations to have an LH:FSH ratio of 1:1. Depending
on the assay methodology used, approximately 6–10 IU of
HCG (equivalent to 50–75 IU LH bioactivity) was found in a
large sample of batches of HMG for the two leading
manufacturers (Stokman, 1993). These urine-derived products
containing equal proportions of LH and FSH activity formed
the mainstay of the burgeoning assisted reproduction market
for the next two decades. Methods of extraction were
continually improved such that more and more impurities
could be excluded from the final preparations. Nonetheless,
the active ingredients (gonadotrophins) still only made up
1–2% of the final product.
As the purity of the products improved, it became apparent
that most women needed FSH rather than LH. Accordingly,
there was a shift from the HMG products that contain FSH and
259
 Articles - New developments in gonadotrophin pharmacology - K Gordon
LH in an approximately 1:1 ratio toward products containing
less and less LH. By the early 1990s, the development of
sophisticated immunopurification and fractionation techniques
allowed the production of highly purified urinary preparations,
some of which are almost totally devoid of LH activity
(Lunenfeld, 2002).
The common α-subunit
Gonadotrophins are composed of a common α-subunit and a
hormone-specific β-subunit (Figure 1). The human α-subunit
is a polypeptide consisting of 92 amino acids, with two N-
linked oligosaccharides joined to asparagine at positions 52
and 78. The α-subunit is common to all three gonadotrophins
and TSH. The human α-subunit is the product of a single gene
(Boothby et al., 1981; Fiddes and Goodman, 1981) and
contains 10 cysteine residues that contribute to the three-
dimensional structure by forming five disulphide bonds.
FSH
The β-subunit of FSH consists of 111 amino acids and has 6
disulphide bonds between its 12 cysteine residues. FSH β-
subunit also has two N-linked glycosylation sites, at positions
7 and 24. The oligosaccharide structures on FSH are highly
variable, with mostly dibranched structures but also tri- or
tetrabranched structures (Ulloa-Aguirre et al., 2001). Each
branch of the oligosaccharides usually terminates in a
negatively charged sialic acid residue or (less often) a
negatively charged sulphate residue. The more sialic acid
residues that terminate oligosaccharide branches, the more
negative the charge (Ulloa-Aguirre and Timossi, 2000).
260 Figure 1. Structure and characteristics of common α- and β-subunits of the human gonadotrophin hormone family.
Human FSH circulates as a mixture of isoforms that can be
separated on the basis of charge (Ulloa-Aguirre and Timossi,
2000). The number and relative abundance of the various
isoforms depend on the source of the sample and change with
the endocrine status and/or stage of the menstrual cycle
(Zambrano et al., 1995). The FSH obtained by recombinant
techniques has slightly more of the higher pH forms than does
the FSH found in HMG. There are currently two commercially
available recombinant FSH products, which, although
produced using slightly different cell lines, are identical in
their amino acid sequences. The isoelectric point (pI) of both
recombinant FSH products is much more acidic than that for
pituitary FSH; hence, all of the commercial preparations have
a longer half-life than that of the native pituitary form.
Horsman et al. (2000) recently compared the charge
distributions of the two commercially available rFSH
preparations and found very little difference, with both
products having essentially the same median pI, between 4.26
and 4.5, and with most of the bulk material fractionated
between 4 and 5 (66.0 ± 1.8% for follitropin-α (Gonal-F®,
Serono International SA, Geneva, Switzerland) and 72.0 ±
1.8% for follitropin-β, Puregon®, NV Organon, Oss, The
Netherlands). Minor differences in charge at extremes of pI
were noted, with Gonal-F® having slightly more of the low pH
forms. However, they do not expect these minor in vitro
differences to produce differences in vivo.
These variations result in an isoform spectrum with isoelectric
points ranging from 3 to 6. The various isoforms differ in their
biological activity, with the more acidic isoforms showing
lower receptor binding activity and lower in-vitro biological
activity in certain assays than do the less-acidic isoforms
 Articles - New developments in gonadotrophin pharmacology - K Gordon
(Ulloa-Aguirre et al., 1995). The impact of the carbohydrate
component of FSH is threefold: first, it contributes to the
serum half-life; second, it impacts proper folding and secretion
at the cellular level; and third, at the target cell level it impacts
on the binding and signal induction mechanisms (Ulloa-
Aguirre et al., 1999). However, it is still unclear whether this
difference in biological responses relates to changes in
association/dissociation or whether binding of different
isoforms to the receptor produces different activations.
Importantly, the isoforms also differ in their metabolic
clearance rate: circulatory half-life is longer for the more
acidic than for the less-acidic isoforms. Thus, the net
biological effect in vivo is a balance between the biological
response and the clearance. However, the relationship between
charge and in-vivo bioactivity is complex.
LH
The β-subunit of LH comprises 121 amino acids with an N-
linked oligosaccharide attached to asparagine at position 30
(Boime et al., 1999). The N-linked oligosaccharides on LH are
mostly sulphated rather than sialylated. This requires N-
acetylgalactosamine (instead of the galactose seen in sialylated
chains) at the penultimate position of the oligosaccharide
chain. Accordingly, it is no surprise that N-acetylgalactosamine
transferase is present in the pituitary but not in the placenta.
There is substantial similarity between LH and CG, with 85%
sequence identity in the first 114 amino acids. This fits with the
fact that both hormones interact with the same receptor, the
LH/HCG receptor.
Figure 2. Structure of FSH-CTP molecule.
Human LH is also present as a heterogeneous mixture of
isoforms, with different patterns present in isolates from the
different sources. Pituitary LH bears oligosaccharides ending
in sulphated branches, whereas recombinant LH lacks sulphate
residues. The synthesis of sulphated structures requires the
consecutive action of GalNAc transferase and
sulphotransferase, which catalyse the addition of the sulphate
group. Chinese hamster ovary (CHO) cells, the main source of
this (commercial) compound, do not express
sulphotransferases. Therefore, human recombinant LH is
mainly sialylated (Amoresano et al., 1996).
A recent report (Talbot et al., 1996) compared the isoform
pattern of two recombinant luteinizing hormone (rLH)
preparations and found that both rLH preparations had LH
activity throughout a broader range of pH than does native
pituitary LH. Pituitary LH was tightly focused in the pI range
of 5.75–4.01, whereas the rLH preparations had about 50–60%
of their activity at pI 5–6. Initial and terminal half-lives were
0.8 and 11 h for rLH and 0.6 and 10 h for pituitary LH (Porchet
et al., 1995). During the follicular phase, LH stimulates theca
cells in the ovaries to secrete androgens, which granulosa cells
use as the substrate to produce oestradiol, thus supporting
FSH-induced follicular development. At mid-cycle, high
levels of LH trigger final oocyte maturation, ovulation, and
corpus luteum formation. After ovulation, LH stimulates the
corpus luteum’s progesterone production, by increasing the
conversion of cholesterol to pregnenolone.
261
 Articles - New developments in gonadotrophin pharmacology - K Gordon
HCG
The β-subunit of HCG is composed of 145 amino acids. HCG
shows substantial structural similarity with human LH.
Furthermore, the two hormones bind to the same receptors.
HCG is distinguished from the β-subunit of LH by the
presence of a carboxy-terminal peptide (CTP) with four O-
linked carbohydrate chains. This extension of the β-subunit of
HCG is believed to produce the prolonged half-life of HCG
compared with that for LH (Boime et al., 1999). Circulating
levels of HCG first become detectable a few days prior to the
missed period, rise rapidly during the first 2 weeks of
pregnancy, and reach maximal levels by week 10–12 of
gestation. Thereafter, levels of HCG slowly decline but remain
detectable until some time after delivery. The only undisputed
role for HCG is maintenance of the corpus luteum, but it may
also contribute to the development of fetal gonadal and adrenal
axes. However, HCG exhibits LH activity and has been used
for years as a surrogate for LH. Also, most urine-derived LH-
containing products contain some HCG.
Source materials for
gonadotrophins
The initial preparations of gonadotrophins were extracted from
pituitary glands of sheep and pigs in the late 1940s. Soon
thereafter, work started on developing a human preparation. In
1958, Carl Gemzell, Egon Diczfalusy, and Gunnar Tillinger
published a report on the ovarian response of seven
amenorrhoeic women treated with a partially purified
preparation of FSH obtained from human pituitaries collected
at autopsy (Gemzell et al., 1958). The treatment proved to be
very effective, but the use of pituitaries from human cadavers
as source material was stopped because of the risk of
iatrogenic transmission of disease. Nevertheless, the practice
of collecting human source material (urine) and harvesting
gonadotrophins continues today.
Urine from menopausal women is the source material for
HMG and the other more ‘purified’ urinary FSH preparations.
It takes approximately 2 l of urine to prepare one 75-IU vial
and upwards of 100 l to complete an average ovarian
stimulation cycle. Hence, an enormous quantity of urine must
be collected to provide the raw material needed for the
production of urinary preparations. Urine from pregnant
women is still collected to be used as the source material for
most of the HCG preparations. Very recently, a recombinant
preparation of HCG has become commercially available with
pharmacokinetics and pharmacodynamics very similar to
those of urine-derived HCG (Trinchard-Lugan, 2002).
The second source of commercial gonadotrophin preparations
is genetically engineered mammalian cell lines that produce
the desired hormone. Recombinant DNA-derived
pharmaceuticals represent a relatively new class of therapeutic
agents. The first of these products (HGH and proinsulin) are
relatively simple polypeptides and were produced using non-
mammalian cell lines (Escherichia coli) (Gesundheit and
Alexander, 1995). The production of glycoproteins presented a
challenge because of the need for proper post-translational
modification (glycosylation) of the protein backbones.
262 However, that challenge has been met: LH, FSH, and HCG
from genetically engineered cell lines are all now
commercially available.
One of the hopes for these new recombinant products was that
they should eliminate problems associated with maintaining
consistency of the quality and quantity of gonadotrophins from
urinary sources. Most would agree that this has proven to be
the case. The various gonadotrophin products are seen in
Table 1.
The future
In future research, attention will probably focus mostly on
modifications of FSH, because FSH is the primary hormone
involved in the development of follicles. With the advent of
recombinant DNA methodologies combined with site-directed
mutagenesis, a variety of structural modifications becomes
possible.
Some clinical researchers have expressed a desire to have three
different preparations of FSH with three different half-lives
(e.g. 6, 12, and 24 h) in order to allow the maximum flexibility
for stimulation of follicle development (Baird, 2001). It is also
anticipated that long-acting formulations would reduce the
number of injections required as part of a stimulation cycle and
that short-acting formulations might reduce the risk of ovarian
hyperstimulation and allow more fine-tuning of ovulation
induction or ovarian stimulation regimens.
FSH-CTP
In 1992, Fares et al. reported that a new follitropin with a
longer half-life had been created by recombinant technology.
The half-life of HCG is longer than that of LH because HCG
has more amino acid and carbohydrate residues at the carboxyl
terminal. The longer half-life of the designer FSH was created
by clipping the end off the β-FSH gene and splicing on the end
of the HCG gene. The new gene was then transfected into
Chinese hamster ovary cells, which then produce the resulting
chimaeric protein FSH-CTP. This product is then extracted
from the medium and purified. FSH-CTP is also known as Org
36286 and by the generic name corifollitropin alfa (Figure 2).
Preclinical pharmacokinetic studies in dogs demonstrated the
expected prolonged half-life for the FSH-CTP, and it was
shown that the compound is bioactive in the classic
Steelman–Pohley assay. A phase I clinical study in 13
hypogonadotrophic hypogonadal (HH) male subjects has been
completed (Bouloux et al., 2001). Since the three-dimensional
structure of this new compound is foreign to humans, the
possible antigenicity of FSH-CTP was of primary concern.
Twelve of the subjects received four 15-µg injections of FSH-
CTP, with an interval of roughly 4 weeks between injections;
one subject received two injections 6 weeks apart. No drug-
related serious adverse events were noted and, importantly, no
antibodies against the new compound were detected. The half-
life of FSH-CTP was about 95 h, or 2–3 times as long as that
of recombinant FSH (Puregon®).
A second trial was then initiated to study the
pharmacokinetics, pharmacodynamics, and safety of Org
36286 after a single subcutaneous injection in 24 healthy
women whose pituitary function was suppressed by 3 weeks of
 Articles - New developments in gonadotrophin pharmacology - K Gordon

Table 1. Summary of the various gonadotrophin products.

Gonadotrophin
products

Product Common trade names Composition Source Route of administration

(manufacturers)
HMG products

Menotropins Humegon 1:1 FSH:LH Purified urinary s.c. or i.m.a

(Organon),
Pergonal (Serono),
Repronex/Menogon (Ferring)
Human menopausal Repronex HP/ 1:1 FSH:LH Highly purified s.c. or i.m.
gonadotrophin Menopur (Ferring) urinary
HCG products

Chorionic A.P.L. (Wyeth-Ayerst), Urinary/highly i.m.

gonadotrophin Novarel (Ferring), purified urinary
Pregnyl (Organon),
Profasi (Serono)
Choriogonadotrophin Ovidrel/Ovitrelle (Serono) Recombinant s.c.
alfa
FSH products

Urofollitropin Metrodin (Serono) Purified urinary i.m.

Urofollitropin Fertinex/Metrodin Highly purified s.c.
HP (Serono), urinary
Bravelle (Ferring)
Follitropin alfa Gonal-F (Serono) Recombinant s.c.
Follitropin beta Puregon/Follistim Recombinant s.c. or i.m.
(Organon)
LH products

Lutropin alfa Luveris (Serono) Recombinant s.c.

aHumegon and Pergonal IM only.

pretreatment with Lyndiol® (50 µg ethinyl oestradiol and 2.5
mg lynestrenol per tablet) (Duijkers et al., 2002). The women
were to continue to take Lyndiol® for up to 3 more weeks after
the Org 36286 injection. Subjects were to be admitted to the
clinic in the evening before day 1 (i.e. the evening before
‘injection day’). Subjects were to stay in the clinic until after
the 36-h sample (on day 2). Serum samples were collected at
various time points for later determination of Org 36286, LH,
inhibin-B, and oestradiol levels.
The women were divided into three groups of eight. The
women in group 1 were to receive a 30 µg dose of Org 36286.
Their responses determined the dose levels for the other
groups. If none of the women in group 1 showed a follicular
response, the Org 36286 dose would be 60 µg for group 2 and
120 µg for group 3. If some but not more than half of the
women in group 1 showed a follicular response, group 2 would
receive 15 µg and group 3 would receive 60 µg. If more than
half of the women in group 1 responded, group 2 would
receive 7.5 µg and group 3 would receive 15 µg.
Initially, three groups of eight women each were treated with
30, 15, and 60 µg, respectively, of Org 36286. However, upon
examination of the data it was concluded that insufficient
response had been seen, so a fourth group consisting of four
subjects from the first group and two from each of the other
two groups were restudied and treated with a dose of 120 µg.
The median maximum numbers of follicles (diameter ≥5 mm)
were 1.0, 1.0, 15.0, and 27.0 follicles (both ovaries added
together) in the 15, 30, 60 and 120 µg groups respectively. The
median day on which the maximum number of follicles was
seen ranged from day 5 to day 9. During the whole study
interval, serum LH levels were very low, thus confirming full
suppression of endogenous gonadotrophins by Lyndiol. The
variety of follicular responses observed with the single
injection of FSH-CTP was similar to that observed with 7 days
of subcutaneous recombinant FSH in a study by Voortman and
colleagues (2000).
Two phase II studies have now been initiated, examining 263
 Articles - New developments in gonadotrophin pharmacology - K Gordon
follicular response in women seeking to become pregnant. The
first study is in women undergoing ovulation induction, and
the second is in women undergoing ovarian stimulation for
IVF/intracytoplasmic sperm injection.
Deglycosylated variants
Complete deglycosylation of HCG results in a compound that
can bind to the LH receptor but can no longer effect the
ordinary biological response.
Single-chain variants
The normal heterodimeric glycoprotein hormones retain their
properties when linked in a single chain. García-Campayo et
al. (1997) reported designing stable biologically active
recombinant LH analogues wherein the carboxy terminus of
the β-subunit is fused to the amino terminus of the α-subunit
through a linker sequence. Furthermore, in the presence of this
linker, the single-chain analogue is secreted efficiently by
transfected Chinese hamster ovary cells and appears stable.
Conclusions
The application of recombinant technology to reproductive
medicine has already led to a number of advances. It is now
possible to design gonadotrophin molecules that mix and
match residues from one gonadotrophin to another, creating
entirely new gonadotrophins. It is anticipated that the
application of these new technologies will lead to more precise
control of exogenous ovarian stimulation (Ulloa-Aguirre and
Timossi, 2001).
References
Amoresano A, Siciliano R, Orru S et al. 1996 Structural
characteristics of human recombinant glycohormones follitropin,
butropin, and choriogonadotrophins expressed in Chinese hamster
ovary cells. European Journal of Biochemistry 242, 608–618.
Baird DT 2001 Is there a place for different isoforms of FSH in
clinical medicine? IV. The clinician’s point of view. Human
Reproduction 16, 1316–1318.
Boime I, Ben-Menahem D, Olijve W 1999 Studies of recombinant
gonadotrophins: intersection of basic science and therapeutics. In:
Fauser BCJM (ed.) Molecular Biology in Reproductive Medicine.
Parthenon Publications, New York, pp. 147–164.
Boothby M, Ruddon RW, Anderson C et al. 1981 A single
gonadotrophin α-subunit gene in normal tissue and tumor-derived
cell lines. Journal of Biological Chemistry 256, 5121–5127.
Bouloux PM, Handelsman DJ, Jockenhovel F et al. 2001 First
human exposure to FSH-CTP in hypogonadotrophic hypogonadal
males. Human Reproduction 16, 1592–1597.
Duijkers IJM, Klipping C, Boerrigter PJ et al. 2002 Single dose
pharmacokinetics and effects on follicular growth and serum
hormones of a long-acting recombinant FSH preparation (FSH-
CTP) in healthy pituitary suppressed females. Human
Reproduction 17, 1987–1993.
Fares FA, Suganuma N, Nishimori K et al. 1992 Design of a long-
acting follitropin agonist by fusing the C-terminal sequence of the
chorionic gonadotrophin β subunit to the follitropin β subunit.
Proceedings of the National Academy of Sciences of the USA 89,
4304–4308.
Fevold HL, Hisaw FL, Leonard SL 1931 The gonad stimulating and
luteinizing hormones of the anterior lobe of the hypophesis.
American Journal of Physiology 97, 291–301.
Fiddes JC, Goodman HM 1981 The gene encoding the common α
264
subunit of the four human glycoprotein hormones. Journal of
Molecular and Applied Genetics 1, 3–18.
García-Campayo V, Sato A, Hirsch B et al. 1997 Design of stable
biologically active recombinant lutropin analogs. Nature
Biotechnology 15, 663–667.
Gemzell CA, Diczfalusy E, Tillinger G 1958 Clinical effect of
human pituitary follicle-stimulating hormone (FSH). Journal of
Clinical Endocrinology and Metabolism 18, 1333–1348.
Gesundheit N, Alexander JK 1995 Endocrine therapy with
recombinant hormones and growth factors. In: Weintraub BD
(ed.) Molecular Endocrinology: Basic Concepts and Clinical
Correlations. Raven Press, New York, pp. 491–507.
Horsman G, Talbot JA, McLoughlin JD et al. 2000 A biological,
immunological and physico-chemical comparison of the current
clinical batches of the recombinant FSH preparations Gonal-F
and Puregon. Human Reproduction 15, 1898–1902.
Lunenfeld B 2002 Development of gonadotrophins for clinical use.
Reproductive BioMedicine Online 4 (suppl. 1), 11–17.
Pierce JG, Parsons TF 1981 Glycoprotein hormones: structure and
function. Annual Review of Biochemistry 50, 465–495.
Porchet HC, Le Cotonnec J-Y, Neuteboom B et al. 1995
Pharmacokinetics of recombinant human luteinizing hormone
after intravenous, intramuscular, and subcutaneous administration
in monkeys and comparison with intravenous administration of
pituitary human luteinizing hormone. Journal of Clinical
Endocrinology and Metabolism 80, 667–673.
Smith PE 1926 Hastening development of female genital system by
daily homoplastic pituitary transplants. Proceedings of the
Society for Experimental Biology and Medicine 24, 131–132.
Stokman PGW, de Leeuw R, van den Wijngaard HAGW et al. 1993
Human chorionic gonadotropin in commercial human
menopausal gonadotropin preparations. Fertility and Sterility 60,
175–178.
Talbot JA, Mitchell R, Hoy AM et al. 1996 Recombinant human
luteinizing hormone: a partial physicochemical, biological and
immunological characterization. Molecular Human Reproduction
2, 799–806.
Trinchard-Lugan I, Khan A, Porchet HC, Munafo A 2002
Pharmacokinetics and pharmcodynamics of recombinant human
chorionic gonadotrophin in healthy male and female volunteers.
Reproductive BioMedicine Online 4, 106–115.
Ulloa-Aguirre A, Timossi C 2000 Biochemical and functional aspects
of gonadotrophin-releasing hormone and gonadotrophins
Reproductive BioMedicine Online 1, 48–62.
Ulloa-Aguirre A, Midgley RA, Beitins IZ, Padmanabhan V 1995
Follicle-stimulating isohormones: characterization and
physiological relevance. Endocrine Reviews 16, 765–787.
Ulloa-Aguirre A, Timossi C, Damian-Matsumura P, Dias JA 1999
Role of glycosylation in function of follicle-stimulating hormone.
Endocrine 11, 205–215.
Ulloa-Aguirre A, Maldonado A, Damian-Matsumura PA, Timossi C
2001 Endocrine regulation of gonadotrophin glycosylation.
Archives of Medical Research 32, 520–532.
Voortman G, Mannaerts BMJL, Huisman JAM 2000 A dose
proportionality study of subcutaneously and intramuscularly
administered recombinant human follicle stimulating hormone
(Follistim/Puregon) in healthy female volunteers. Fertility and
Sterility 73, 1187–1193.
Zambrano E, Olivares A, Mendez JP et al. 1995 Dynamics of basal
and gonadotropin-releasing hormone–releasable serum follicle-
stimulating hormone charge isoform distribution throughout the
human menstrual cycle. Journal of Clinical Endocrinology and
Metabolism 80, 1647–1656.
Paper based on contribution presented at the Second Expert Organon
meeting in Rottach-Egern, Germany, March 2002.