0013-7227/91/1295-2277$03.00/0 Endocrinology Copyright © 1991 by The Endocrine Society
Vol. 129, No. 5 Printed in U.S.A.
The Early Days of Steroid RadioimmiLinoassays at the Worcester Foundation In the late 1960s, the Worcester Foundation for Experimental Biology in Shrewsbury, Massachusetts continued to maintain its position as a veritable "Mecca" of steroid research. The faculty included Drs. A. and H. Brodie, Burstein, Caspi, Chang, Lloyd, McCracken, Peron, Romanoff, S. and J. Tait, Townsley, Williams, and others. Postdoctoral training programs in Steroid Biochemistry and in Reproductive Biology were thriving, a distillation plant was producing "in house" organic solvents of chromatographic grade, and TLC, Bush I, and Bush II were parts of the vocabulary. The "double isotope" and "protein binding" assays were in routine use for quantitation of steroid hormones in biological fluids. At the same time, Dr. Guy Abraham was laboring in a one-person laboratory on an idea of devising a radioimmunoassay for estradiol. Unless I am mistaken, most of the biochemists and "mainstream" steroid workers were giving this project a probability of success which was not significantly different from zero. However, Guy was about to prove everyone wrong with his success in
developing a solid-phase radioimmunoassay system for estradiol-170 (1). At about the same time, a group of young scientists associated with the training programs was busy immunizing animals with various steroid conjugates in an attempt to interfere with hormone action by active and passive immunization and to develop radioimmunoassays. This group consisted of Drs. Burton Caldwell, Rex Scaramuzzi, Steven Tillson, and Ian Thorneycroft. Their efforts were singularly successful with a string of exciting papers appearing in various journals, and radioimmunoassays for progesterone, testosterone, and androstenedione becoming a reality in rapid succession. This was soon followed by a report at an opening session of The 1971 Endocrine Society Meeting of successful development of RIA for prostaglandin F2a (2). The pace of progress in this group was rapid and publication of the various methods often lagged behind. Also, there seemed Received July 10,1991. "Remembrance," articles discuss people and events as remembered by the author. The opinion(s) expressed are solely those of the writer and do not reflect the view of the Journal or The Endocrine Society. NOTE: This brief assay is a personal view from an observer of a particular stage of steroid research at one institution. No attempt was made to provide a comprehensive review or to give credit to all those who provided help, financial support, and facilities for these studies.
to be little concern for establishing a written record of who should receive credit for development of these procedures, and exactly when the various assays were first developed. In the meantime, Burt Caldwell, who headed this group, and his colleagues were wonderfully open about their work and not only willingly shared the antisera and assay protocols with whoever was interested, but publicized availability of the new assays and offered assistance in establishing them in other laboratories. As a result, some of these procedures were first described in publications from laboratories that did not participate in their development [e.g. testosterone RIA (3)]. Similarly, the critical role of Dr. Sumner Burstein who synthesized steroid conjugates (I believe mostly BSA-oximes and hemi-succinates) for these studies, or help provided by Drs. Tom Wittstruck and Ken Williams with magnetic resonance analysis and organic chemistry, would probably not be obvious from reading the literature. Performance of the newly developed radioimmunoassays elicited a mixture of awe and incredulity. Not only were these procedures more sensitive than the previously available methods, but they were much less labor intensive and easier to learn. However, occasional difficulties with blank values or column chromatography tended to reinforce the suspicion that this new "magic" may be too good to be true. Hence, both the developers and the early users of these assays spent much time and energy on testing, validating, and comparing the results to those obtained by established methods. It was felt that chromatographic preparation of the sample was a must and the possibility of doing assays on unextracted serum or plasma was not even considered. Now, nearly 20 years later, this period seems very remote indeed. Radioimmunoassays for steroids are "routine" to the point that many manuscripts limit description of these procedures to quoting a single reference from another laboratory! Obviously the Worcester Foundation was not the only place where steroid radioimmunoassays were being successfully developed. However, I believe that the impact of the procedures developed at this institution on the progress of endocrinology was remarkable. For many years, acknowledgments of Drs. Caldwell, Thorneycroft, and Abraham as providers of antisera appeared in what must have been hundreds of papers and there is little
2277
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 25 November 2015. at 05:24 For personal use only. No other uses without permission. . All rights reserved.
2278
REMEMBRANCE
doubt that a significant proportion of the world's literature dealing with reproductive endocrinology in the 1970s reported results of studies performed with the use of these reagents. I had a supply of Burt Caldwell's antiserum to testosterone until 1985! In the present era of commercial RIA labs, easy-to-use kits, and sophisticated mathematical analysis of hormone levels measured in hundreds of serially collected samples, it is interesting to realize that as recently as 20 years ago measurements of steroid hormones were far from routine and it was exciting to be able to measure steroids in the amount of blood that could be collected from a mouse or a juvenile rat.
Endo• 1991 Voll29«No5
Andrzej Bartke Professor and Chairman Department of Physiology Southern Illinois University School of Medicine
References 1. Abraham GA 1969 Solid-phase radioimmunoassay of estradiol-17,s. J Clin Endocrinol Metab 29:866-870 2. Caldwell BV, Brock WA, Burstein S, Speroff L, Radioimmunoassay of prostaglandin F2a. Program of the 53rd Annual Meeting of The Endocrine Society, San Francisco, CA, A-46, 1971 (Abstract 8) 3. Bartke A, Steele RE, Musto N, Caldwell BW 1973 Fluctuations in plasma testosterone levels in adult male rats and mice. Endocrinology 91:1223-1228
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 25 November 2015. at 05:24 For personal use only. No other uses without permission. . All rights reserved.
Vol. 129, No. 5 Printed in U.S.A.
The Early Days of Steroid RadioimmiLinoassays at the Worcester Foundation In the late 1960s, the Worcester Foundation for Experimental Biology in Shrewsbury, Massachusetts continued to maintain its position as a veritable "Mecca" of steroid research. The faculty included Drs. A. and H. Brodie, Burstein, Caspi, Chang, Lloyd, McCracken, Peron, Romanoff, S. and J. Tait, Townsley, Williams, and others. Postdoctoral training programs in Steroid Biochemistry and in Reproductive Biology were thriving, a distillation plant was producing "in house" organic solvents of chromatographic grade, and TLC, Bush I, and Bush II were parts of the vocabulary. The "double isotope" and "protein binding" assays were in routine use for quantitation of steroid hormones in biological fluids. At the same time, Dr. Guy Abraham was laboring in a one-person laboratory on an idea of devising a radioimmunoassay for estradiol. Unless I am mistaken, most of the biochemists and "mainstream" steroid workers were giving this project a probability of success which was not significantly different from zero. However, Guy was about to prove everyone wrong with his success in
developing a solid-phase radioimmunoassay system for estradiol-170 (1). At about the same time, a group of young scientists associated with the training programs was busy immunizing animals with various steroid conjugates in an attempt to interfere with hormone action by active and passive immunization and to develop radioimmunoassays. This group consisted of Drs. Burton Caldwell, Rex Scaramuzzi, Steven Tillson, and Ian Thorneycroft. Their efforts were singularly successful with a string of exciting papers appearing in various journals, and radioimmunoassays for progesterone, testosterone, and androstenedione becoming a reality in rapid succession. This was soon followed by a report at an opening session of The 1971 Endocrine Society Meeting of successful development of RIA for prostaglandin F2a (2). The pace of progress in this group was rapid and publication of the various methods often lagged behind. Also, there seemed Received July 10,1991. "Remembrance," articles discuss people and events as remembered by the author. The opinion(s) expressed are solely those of the writer and do not reflect the view of the Journal or The Endocrine Society. NOTE: This brief assay is a personal view from an observer of a particular stage of steroid research at one institution. No attempt was made to provide a comprehensive review or to give credit to all those who provided help, financial support, and facilities for these studies.
to be little concern for establishing a written record of who should receive credit for development of these procedures, and exactly when the various assays were first developed. In the meantime, Burt Caldwell, who headed this group, and his colleagues were wonderfully open about their work and not only willingly shared the antisera and assay protocols with whoever was interested, but publicized availability of the new assays and offered assistance in establishing them in other laboratories. As a result, some of these procedures were first described in publications from laboratories that did not participate in their development [e.g. testosterone RIA (3)]. Similarly, the critical role of Dr. Sumner Burstein who synthesized steroid conjugates (I believe mostly BSA-oximes and hemi-succinates) for these studies, or help provided by Drs. Tom Wittstruck and Ken Williams with magnetic resonance analysis and organic chemistry, would probably not be obvious from reading the literature. Performance of the newly developed radioimmunoassays elicited a mixture of awe and incredulity. Not only were these procedures more sensitive than the previously available methods, but they were much less labor intensive and easier to learn. However, occasional difficulties with blank values or column chromatography tended to reinforce the suspicion that this new "magic" may be too good to be true. Hence, both the developers and the early users of these assays spent much time and energy on testing, validating, and comparing the results to those obtained by established methods. It was felt that chromatographic preparation of the sample was a must and the possibility of doing assays on unextracted serum or plasma was not even considered. Now, nearly 20 years later, this period seems very remote indeed. Radioimmunoassays for steroids are "routine" to the point that many manuscripts limit description of these procedures to quoting a single reference from another laboratory! Obviously the Worcester Foundation was not the only place where steroid radioimmunoassays were being successfully developed. However, I believe that the impact of the procedures developed at this institution on the progress of endocrinology was remarkable. For many years, acknowledgments of Drs. Caldwell, Thorneycroft, and Abraham as providers of antisera appeared in what must have been hundreds of papers and there is little
2277
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 25 November 2015. at 05:24 For personal use only. No other uses without permission. . All rights reserved.
2278
REMEMBRANCE
doubt that a significant proportion of the world's literature dealing with reproductive endocrinology in the 1970s reported results of studies performed with the use of these reagents. I had a supply of Burt Caldwell's antiserum to testosterone until 1985! In the present era of commercial RIA labs, easy-to-use kits, and sophisticated mathematical analysis of hormone levels measured in hundreds of serially collected samples, it is interesting to realize that as recently as 20 years ago measurements of steroid hormones were far from routine and it was exciting to be able to measure steroids in the amount of blood that could be collected from a mouse or a juvenile rat.
Endo• 1991 Voll29«No5
Andrzej Bartke Professor and Chairman Department of Physiology Southern Illinois University School of Medicine
References 1. Abraham GA 1969 Solid-phase radioimmunoassay of estradiol-17,s. J Clin Endocrinol Metab 29:866-870 2. Caldwell BV, Brock WA, Burstein S, Speroff L, Radioimmunoassay of prostaglandin F2a. Program of the 53rd Annual Meeting of The Endocrine Society, San Francisco, CA, A-46, 1971 (Abstract 8) 3. Bartke A, Steele RE, Musto N, Caldwell BW 1973 Fluctuations in plasma testosterone levels in adult male rats and mice. Endocrinology 91:1223-1228
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 25 November 2015. at 05:24 For personal use only. No other uses without permission. . All rights reserved.
