BG1Luc Background Review Document Annexes

ICCVAM Test Method Evaluation Report
The LUMI-CELL
® ER (BG1Luc ER TA) Test Method: An In Vitro Assay for Identifying Human Estrogen Receptor Agonist and Antagonist Activity of Chemicals

NIH Publication No. 11-7850

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Annex A: Submission of XDS’s LUMI-CELL™ ER High-Throughput System for Screening Estrogen-Like Chemicals for Review by ICCVAM

Annex B: ICCVAM/NICEATM Evaluation – BG1Luc ER TA Submission

Annex C: NICEATM Report on the XDS Protocol Standardization Study Agonist and Antagonist Protocols

Annex D: Addendum to NIH Publication No: 03-4503 

Annex E: ICCVAM/NICEATM BG1Luc ER TA – Agonist Protocol

Annex F: ICCVAM/NICEATM BG1Luc ER TA – Antagonist Protocol

Annex G: Materials Relating to Cell Viability

Annex H: ICCVAM/NICEATM BG1Luc4E2 ER TA – Validation Work Plan

Annex I: Substances Used for the Validation of the BG1Luc ER TA Test Method

Annex J: Prism Files for All Validation Study Phases

Graphs ("PZF" files) may be viewed using GraphPad Prism® or Prism® viewer software. The Prism® viewer software is a free download available at the GraphPad website

  • Phase 1 Graphs
    • Agonist Reference Standard [PZF]
    • Antagonist Reference Standard [PZF]
  • Phase 2a Range Finder Testing
    • Phase 2a Agonist Range Finder 1 [PZF]
    • Phase 2a Agonist Range Finder 2 [PZF]
    • Phase 2a Antagonist Range Finder 1 [PZF]
    • Phase 2a Antagonist Range Finder 2 [PZF]
  • Phase 2a Comprehensive Agonist Testing
    • Cross Laboratory Test Substance Comparison [PZF]
    • Bisphenol A (ECVAM) [PZF]
    • Bisphenol A (Hiyoshi) [PZF]
    • Bisphenol A (XDS) [PZF]
    • Bisphenol B (ECVAM) [PZF]
    • Bisphenol B (Hiyoshi) [PZF]
    • Bisphenol B (XDS) [PZF]
    • Corticosterone (ECVAM) [PZF]
    • Corticosterone (Hiyoshi) [PZF]
    • Corticosterone (XDS) [PZF]
    • Diethylstilbestrol (ECVAM) [PZF]
    • Diethylstilbestrol (Hiyoshi) [PZF]
    • Diethylstilbestrol (XDS) [PZF]
  • Phase 2a Comprehensive Antagonist Testing
    • Cross Laboratory Test Substance Comparison [PZF]
    • Dibenzo[a.h]anthracene (ECVAM) [PZF]
    • Dibenzo[a.h]anthracene (Hiyoshi) [PZF]
    • Dibenzo[a.h]anthracene (XDS) [PZF]
    • p-n-nonylphenol (ECVAM) [PZF]
    • p-n-nonylphenol (Hiyoshi) [PZF]
    • p-n-nonylphenol (XDS) [PZF]
    • Progesterone (ECVAM) [PZF]
    • Progesterone (Hiyoshi) [PZF]
    • Progesterone (XDS) [PZF]
    • Tamoxifen (ECVAM) [PZF]
    • Tamoxifen (Hiyoshi) [PZF]
    • Tamoxifen (XDS) [PZF]
  • Phase 2b Range Finder Testing
    • Phase 2b Range Finder (ECVAM) [PZF]
    • Phase 2b Range Finder (Hiyoshi) [PZF]
    • Phase 2b Range Finder (XDS) [PZF]
  • Phase 2b Comprehensive Agonist Testing
    • Cross Laboratory Test Substance Comparison [PZF]
    • 17-α ethinyl estradiol (ECVAM) [PZF]
    • 17-α ethinyl estradiol (Hiyoshi) [PZF]
    • 17-α ethinyl estradiol (XDS) [PZF]
    • Atrazine (ECVAM) [PZF]
    • Atrazine (ECVAM retest) [PZF]
    • Atrazine (Hiyoshi) [PZF]
    • Atrazine (XDS) [PZF]
    • Butylbenzyl phthalate (ECVAM) [PZF]
    • Butylbenzyl phthalate (Hiyoshi) [PZF]
    • Butylbenzyl phthalate (XDS) [PZF]
    • Flavone (ECVAM) [PZF]
    • Flavone (Hiyoshi) [PZF]
    • Flavone (XDS) [PZF]
    • Genistein (ECVAM) [PZF]
    • Genistein (Hiyoshi) [PZF]
    • Genistein (XDS) [PZF]
    • o.p’-DDT (ECVAM) [PZF]
    • o.p’-DDT (Hiyoshi) [PZF]
    • o.p’-DDT (XDS) [PZF]
    • p-n-nonylphenol (ECVAM) [PZF]
    • p-n-nonylphenol (Hiyoshi) [PZF]
    • p-n-nonylphenol (XDS) [PZF]
    • Vinclozolin (ECVAM) [PZF]
    • Vinclozolin (ECVAM retest) [PZF]
    • Vinclozolin (Hiyoshi) [PZF]
    • Vinclozolin (XDS) [PZF]
  • Phase 2b Comprehensive Antagonist Testing
    • Cross Laboratory Test Substance Comparison [PZF]
    • Apigenin (ECVAM) [PZF]
    • Apigenin (Hiyoshi) [PZF]
    • Apigenin (XDS) [PZF]
    • Apigenin (XDS retest) [PZF]
    • Atrazine (ECVAM) [PZF]
    • Atrazine (Hiyoshi) [PZF]
    • Atrazine (XDS) [PZF]
    • Atrazine (XDS retest) [PZF]
    • Butylbenzyl phthalate (ECVAM) [PZF]
    • Butylbenzyl phthalate (Hiyoshi) [PZF]
    • Butylbenzyl phthalate (XDS) [PZF]
    • Corticosterone (ECVAM) [PZF]
    • Corticosterone (Hiyoshi) [PZF]
    • Corticosterone (XDS) [PZF]
    • Flavone (ECVAM) [PZF]
    • Flavone (Hiyoshi) [PZF]
    • Flavone (Hiyoshi retest) [PZF]
    • Flavone (XDS) [PZF]
    • Genistein (ECVAM) [PZF]
    • Genistein (Hiyoshi) [PZF]
    • Genistein (Hiyoshi retest) [PZF]
    • Genistein (XDS) [PZF]
    • Genistein (XDS retest) [PZF]
    • o.p’-DDT (ECVAM) [PZF]
    • o.p’-DDT (Hiyoshi) [PZF]
    • o.p’-DDT (XDS) [PZF]
    • o.p’-DDT (XDS retest) [PZF]
    • Resveratrol (ECVAM) [PZF]
    • Resveratrol (Hiyoshi) [PZF]
    • Resveratrol (XDS) [PZF]
    • Resveratrol (XDS retest) [PZF]
  • Phase 3 Range Finder Testing
    • Phase 3 Agonist Range Finder (ECVAM) [PZF]
    • Phase 3 Agonist Range Finder (Hiyoshi) [PZF]
    • Phase 3 Agonist Range Finder (XDS) [PZF]
    • Phase 3 Antagonist Range Finder (ECVAM) [PZF]
    • Phase 3 Antagonist Range Finder (Hiyoshi) [PZF]
    • Phase 3 Antagonist Range Finder (XDS) [PZF]
  • Phase 3 Comprehensive Agonist Testing
    • 12-O-Tetradecanoylphorbol-13-acetate [PZF]
    • 17-α estradiol [PZF]
    • 17-β estradiol [PZF]
    • 2-sec-butylphenol [PZF]
    • 2,4,5-trichlorophenoxyacetic acid [PZF]
    • 4-androstenedione [PZF]
    • 4-cumylphenol [PZF]
    • 4-hydroxytamoxifen [PZF]
    • 4-tert-octylphenol [PZF]
    • 5a-dihydrotestosterone [PZF]
    • Actinomycin D [PZF]
    • Apigenin [PZF]
    • Clomiphene citrate [PZF]
    • Coumestrol [PZF]
    • Daidzein [PZF]
    • Dexamethasone [PZF]
    • Di-n-butyl phthalate [PZF]
    • Dibenzo[a.h]anthracene [PZF]
    • Dicofol [PZF]
    • Diethylhexyl phthalate [PZF]
    • Estrone [PZF]
    • Ethyl paraben [PZF]
    • Fluoranthene [PZF]
    • Hydroxyflutamide [PZF]
    • Kaempferol [PZF]
    • Kepone [PZF]
    • meso-hexestrol [PZF]
    • Methyl testosterone [PZF]
    • Morin [PZF]
    • Norethynodrel [PZF]
    • p.p’-DDE [PZF]
    • p.p’-methoxychlor [PZF]
    • Phenobarbital [PZF]
    • Phenolphthalin [PZF]
    • Progesterone [PZF]
    • Propylthiouracil [PZF]
    • Raloxifene HCl [PZF]
    • Resveratrol [PZF]
    • Sodium azide [PZF]
    • Tamoxifen [PZF]
    • Testosterone [PZF]
  • Phase 3 Comprehensive Antagonist Testing
    • 12-O-Tetradecanoylphorbol-13-acetate [PZF]
    • 17-α estradiol [PZF]
    • 17-β estradiol [PZF]
    • 17α- ethinyl estradiol [PZF]
    • 2-sec-butylphenol [PZF]
    • 2,4,5-trichlorophenoxyacetic acid [PZF]
    • 4-androstenedione [PZF]
    • 4-cumylphenol [PZF]
    • 4-hydroxytamoxifen [PZF]
    • 4-tert-octylphenol [PZF]
    • 5a-dihydrotestosterone [PZF]
    • Actinomycin D [PZF]
    • Bisphenol A [PZF]
    • Bisphenol B [PZF]
    • Clomiphene citrate [PZF]
    • Coumestrol [PZF]
    • Daidzein [PZF]
    • Dexamethasone [PZF]
    • Di-n-butyl phthalate [PZF]
    • Dicofol [PZF]
    • Diethylhexyl phthalate [PZF]
    • Diethylstilbestrol [PZF]
    • Estrone [PZF]
    • Ethyl paraben [PZF]
    • Fluoranthene [PZF]
    • Hydroxyflutamide [PZF]
    • Kaempferol [PZF]
    • Kepone [PZF]
    • meso-hexestrol [PZF]
    • Methyl testosterone [PZF]
    • Morin [PZF]
    • Norethynodrel [PZF]
    • p.p’-DDE [PZF]
    • p.p’-methoxychlor [PZF]
    • Phenobarbital [PZF]
    • Phenolphthalin [PZF]
    • Propylthiouracil [PZF]
    • Raloxifene HCl [PZF]
    • Sodium azide [PZF]
    • Testosterone [PZF]
    • Vinclozolin [PZF]
  • Phase 4 Range Finder Testing
    • Phase 4 Agonist Range Finder [PZF]
    • Phase 4 Antagonist Range Finder [PZF]
  • Phase 4 Comprehensive Agonist Testing
    • 17β-trenbolone [PZF]
    • 19-nortestosterone [PZF]
    • 2,2’,4,4’-Tetrahydroxy benzophenone [PZF]
    • 4-hydroxy androstenedione [PZF]
    • Ammonium perchlorate [PZF]
    • Apomorphine [PZF]
    • Bicalutamide [PZF]
    • Chrysin [PZF]
    • Cycloheximide [PZF]
    • Cyproterone acetate [PZF]
    • Fenarimol [PZF]
    • Finasteride [PZF]
    • Fluoxymestrone [PZF]
    • Flutamide [PZF]
    • Haloperidol [PZF]
    • Ketoconazole [PZF]
    • L-thyroxine [PZF]
    • Linuron [PZF]
    • Medroxyprogesterone acetate [PZF]
    • Mifepristone [PZF]
    • Nilutamide [PZF]
    • Oxazepam [PZF]
    • Pimozide [PZF]
    • Procymidone [PZF]
    • Reserpine [PZF]
    • Spironolactone [PZF]
  • Phase 4 Comprehensive Antagonist Testing
    • 17β-trenbolone [PZF]
    • 19-nortestosterone [PZF]
    • 4-hydroxy androstenedione [PZF]
    • Ammonium perchlorate [PZF]
    • Apomorphine [PZF]
    • Bicalutamide [PZF]
    • Chrysin [PZF]
    • Cycloheximide [PZF]
    • Cyproterone acetate [PZF]
    • Fenarimol [PZF]
    • Finasteride [PZF]
    • Fluoxymestrone [PZF]
    • Flutamide [PZF]
    • Haloperidol [PZF]
    • Ketoconazole [PZF]
    • L-thyroxine [PZF]
    • Linuron [PZF]
    • Medroxyprogesterone acetate [PZF]
    • Mifepristone [PZF]
    • Nilutamide [PZF]
    • Oxazepam [PZF]
    • Pimozide [PZF]
    • Procymidone [PZF]
    • Reserpine [PZF]
    • Spironolactone [PZF]

Annex K: Experimental Details

Annex L: Within-Plate DMSO Control Values for BG1Luc ER TA Agonist and Antagonist Assays

Annex M: Plate Redesign and Compilation of Historical Database

Annex N: Literature Update Files