Discrimination of recombinant from natural human growth hormone using DNA aptamers

1 Apr 2011

Discrimination of recombinant from natural human growth hormone using DNA aptamers / John G. Bruno, Maria P. Carrillo, Taylor Phillips, Allison Edge. - (Journal of Biomolecular Techniques 22 (2011) 1 (April) ; p. 27-36)

  • PMID: 21455479
  • PMCID: PMC3059541

Abstract

Detection of athletes who use synthetic human growth hormone (hGH; or somatotropin) to enhance physical strength and obtain an advantage in competitive sports is a formidable problem, as rhGH is virtually identical to the natural pituitary hormone. However, some post-translational and other modifications have been documented by chromatographic separation and mass spectrometry (MS) in a small percentage of rhGH. In the present work, development of DNA aptamers against research-grade rhGH and natural hGH with adsorption of the rhGH aptamers against natural hGH was shown to produce a small family of aptamer sequences that bound consistently with greater affinity to rhGH over a low nanogram-to-microgram range in ELISA-like microplate assays. This collection of rhGH discriminatory aptamer sequences shared some short sequence segments and secondary structural features. The top rhGH discriminatory aptamers also appeared to cross-react with human myoglobin and BSA but not with bone collagen peptides and an unrelated viral envelope peptide. The cross-reactivity results suggested several strings of up to five consecutive amino acids that might serve as common epitopes for aptamer binding. SDS-PAGE revealed that the rhGH existed largely as a 45-kDa dimer, and the natural hGH was almost exclusively monomeric. The existence of the rhGH dimer suggests that a discontinuous "bridge" epitope may exist on the rhGH, which spans the subunits, thereby accounting somewhat for the difference in detection. Overall, these results suggest that aptamers might be useful for routine, presumptive laboratory screening to identify athletes who are potentially cheating by administration of rhGH.

Detection of GH abuse in sport: Past, present and future

30 May 2009

Detection of GH abuse in sport: Past, present and future / Osquel Barroso, Patrick Schamasch, Olivier Rabin. - (Growth Hormone & IGF Research 19 (2009) 4 (August) ; p. 369-374)

  • PMID: 19482501
  • DOI: 10.1016/j.ghir.2009.04.021

Abstract:

Due to its considered performance enhancing effects, human growth hormone (hGH) is abused as a doping agent in sport. Its misuse also carries potentially serious side effects to a person’s health. Consequently, hGH and its releasing factors are prohibited in sport, as established in the Prohibited List which is updated and published yearly by the World Anti-Doping Agency (WADA). In order to fight the menace that hGH doping poses to the spirit of sport and to the health of athletes, the sport movement and the anti-doping authorities, initially led by the International Olympic Committee (IOC) and later by WADA, have put substantial efforts into developing tests for its detection. Currently, a primary analytical approach, the isoform differential immunoassay, has been implemented in WADA-accredited laboratories. In parallel, a second, indirect approach for the detection of hGH abuse, based on the quantification of hGH-associated biological markers, has been developed. The final aim is to combine both methodologies to improve the sensitivity and expand the time window to detect doping with hGH. In addition, novel analytical procedures, based on proteomic and genomic technologies as well as the use of mass spectrometry-based methods of detection, are being investigated for future application in hGH anti-doping tests.

CAS 2003_A_484 Kicker Vencill vs USADA - Interim award

18 Nov 2003

CAS 2003/A/484 Kicker Vencill vs USADA - Interim award

The Court of Arbitration decides on 18 November 2002:

1. The Jurisdiction oif CAS is affirmed;

2. Ibe appeal filed by mr. Vencill on 14 July 2003 is dismissed;

CAS 2003_A_484 Kicker Vencill vs USADA - Final award

11 Mar 2004

CAS 2003/A/484 Kicker Vencill vs USADA

Related documents:

  • AAA No. 30 190 00291 03 USADA vs Kicker Vencill
    July 24, 2003
  • CAS 2003_A_484 Kicker Vencill vs USADA - Interim award
    November 18, 2003
  • USADA - Supplement 411 - Kicker Vencill, Introduction Video
    May 24, 2012


On 23 June 2003 the North American Court of Arbitration (NACAS) decided to impose a 4 year period of ineligibility on the American swimmer Kicker Vencill after his A and B samples tested positive for the prohibited substance 19-norandrosterone (Nandrolone).

Hereafter in July 2003 the Athlete appealed the NACAS decision with the Court of Arbitration for Sport (CAS). The Athlete requested for a reduced sanction on the basis of No Significant Fault or Negligence.

The Athlete argued a number of issues in support of his appeal, ranging from:

  • questions concerning the chain of custody of his sample;
  • alleged violations of his right to be present for the testing of his B sample;
  • supposed inaccuracies in the results reported by the UCLA Lab; and
  • allegations to the effect that the low concentration of 19-norandrosterone found in the athlete's sample is consistent with endogenous production as opposed to exogenoμs administration or ingestion of a prohibited substance.

The Panel finds that there is no question that the Athlete is guilty of committing an anti-doping rule violation and he failed to establish that the chain of custody of his sample was anything other than intact. Further the Panel concludes that the laboratory analysis was correctly conducted, the Athlete’s samples had not deteriorated or been contaminated and the proper laboratory procedures had been followed.

The Panel accepts that the violation was not intentional and that laboratory analysis revealed that the supplement in question was contaminated. However the Athlete showed also a total disregard of his positive duty to ensure that no prohibited substance enters his body.

As a result the Panel holds that the Athlete’s Fault or Negligence in the circumstances is exceptionally significant in relation to the doping violation.

Therefore the of Court of Arbitration for Sport decides on 11 March 2004 that:

1.) The jurisriction of the CAS is affirmed;

2.) The appeal filed by Mr. Vencill on 14 July 2003 is dismissed;

3.) Save for the applicable period of ineligibility as specified in paragraph 4 below, the decision in this matter issued by the North American Court of Arbitration for Sport Panel dated 23 June 2003 is upheld;

4.) Kicker Vencill shall be declared ineligible for competition for two years commencing as of 22 May 2003:

5.) The Court Office fee of CHF 500 already paid by Mr. Vencill shall be retained by the CAS;

6.) Each party shall bear its own costs.

A false start in the race against doping in sport: concerns with cycling's biological passport

1 Nov 2011

A false start in the race against doping in sport: concerns with cycling's biological passport / Nicolas Hailey. - (Duke law journal 61 (2011) 2 (November) ; 393-432).

  • PMID: 22069854

Abstract:

Professional cycling has suffered from a number of doping scandals. The sport's governing bodies have responded by implementing an aggressive new antidoping program known as the biological passport. Cycling's biological passport marks a departure from traditional antidoping efforts, which have focused on directly detecting prohibited substances in a cyclist's system. Instead, the biological passport tracks biological variables in a cyclist's blood and urine over time, monitoring for fluctuations that are thought to indirectly reveal the effects of doping. Although this method of indirect detection is promising, it also raises serious legal and scientific concerns. Since its introduction, the cycling community has debated the reliability of indirect biological-passport evidence and the clarity, consistency, and transparency of its use in proving doping violations. Such uncertainty undermines the legitimacy of finding cyclists guilty of doping based on this indirect evidence alone. Antidoping authorities should address these important concerns before continuing to pursue doping sanctions against cyclists solely on the basis of their biological passports.

Sports doping : Racing just to keep up.

15 Jul 2011

Sports doping : Racing just to keep up / Ewen Callaway. - (Nature 475 (2011) 7356 (21 July) ; p. 283-285)

  • PMID: 21776058.
  • DOI: 10.1038/475283a

Anti-doping researchers are looking for new ways to catch cheaters. Can a biological passport help to save the sport?

Gene doping [2006]

1 Apr 2006

Genetische doping / H.J. Haisma, Olivier de Hon. - (Nederlands Tijdschrift voor Klinische Chemie en Laboratoriumgeneeskunde 31 (2006) 1 : p. 261-263)

Genetische doping zal zeer waarschijnlijk binnen 5 jaar zijn intrede doen in de sportwereld. Bepaalde genen kunnen de sportieve prestatie immers verbeteren. Deze genen worden thans onderzocht in klinische studies voor de behandeling van ziektes. Het niet-therapeutische en ongecontroleerde gebruik van gendoping door sporters kan resulteren in gezondheidsschade.
Preventieve maatregelen om het gebruik van gendoping te voorkomen zijn dus noodzakelijk. Een uitvoerig voorlichtingsprogramma t.b.v. de sportbegeleiders en de sporters zelf, een evaluatie van de huidige regelgeving en de ontwikkeling van een geavanceerde detectiemethode op basis van proteomische technieken lijken de meest veelbelovende preventieve maatregelen.

Gene doping : of mice and men.

20 Jan 2009

Gene doping : of mice and men / Hassan M.E. Azzazya, Mai M.H. Mansoura, Robert H. Christenson. - (Clinical Biochemistry 42 (2009) 6 (April) ; p. 435-441)

  • PMID: 19272337.
  • DOI: 10.1016/j.clinbiochem.2009.01.001


Abstract

Gene doping is the newest threat to the spirit of fair play in sports. Its concept stemmed out from legitimate gene therapy trials, but anti-doping authorities fear that they now may be facing a form of doping that is virtually undetectable and extremely appealing to athletes. This paper presents studies that generated mouse models with outstanding physical performance, by manipulating genes such as insulin-like growth factor 1 (IGF-1) or phosphoenolpyruvate carboxykinase (PEPCK), which are likely to be targeted for gene doping. The potential transition from super mice to super athletes will also be discussed, in addition to possible strategies for detection of gene doping.

Ethics : Gene doping and sport

5 Feb 2010

Ethics : Gene doping and sport / Theodore Friedmann, Olivier Rabin, Mark S. Frankel. - (Science 327 (2010) 5966 (5 February) ; p. 647-648)

  • PMID: 20133558.
  • DOI: 10.1126/science.1177801

Blood doping and its detection

1 Sep 2011

Blood doping and its detection / Wolfgang Jelkmann, Carsten Lundby. - (Blood 118 (2011) 9 (1 September) p. 2395-2404).

  • PMID: 21652677.
  • DOI: 10.1182/blood-2011-02-303271

Abstract:

Hemoglobin mass is a key factor for maximal exercise capacity. Some athletes apply prohibited techniques and substances with intent to increase hemoglobin mass and physical performance, and this is often difficult to prove directly. Autologous red blood cell transfusion cannot be traced on reinfusion, and also recombinant erythropoietic proteins are detectable only within a certain timeframe. Novel erythropoietic substances, such as mimetics of erythropoietin (Epo) and activators of the Epo gene, may soon enter the sports scene. In addition, Epo gene transfer maneuvers are imaginable. Effective since December 2009, the World Anti-Doping Agency has therefore implemented "Athlete Biologic Passport Operating Guidelines," which are based on the monitoring of several parameters for mature red blood cells and reticulocytes. Blood doping may be assumed, when these parameters change in a nonphysiologic way. Hematologists should be familiar with blood doping practices as they may play an important role in evaluating blood profiles of athletes with respect to manipulations, as contrasted with the established diagnosis of clinical disorders and genetic variations.

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