laboratory of immunobiochemistry allergenic products advisory committee, april 8, 2003

Laboratory of Immunobiochemistry

Allergenic Products Advisory Committee, April 8, 2003

Allergenic Products Advisory Committee, April 8, 2003

Lab overview Staffing Lot release Reference replacement

Operational issues Sheep serum replacement issues ISO certification report

Research/regulatory update Endotoxin studies Cockroach antigens and antibodies

Lab overview Staffing Lot release Reference replacement

Principal Investigators Jay E. Slater, MD, Lab Chief – Supervisory

Medical Officer (4) Ronald Rabin, MD - Senior Staff Fellow (2)

Post Doctoral Fellows

Jonny Finlay, PhD - IRTA (2) Bo Chi, MD - Visiting Associate (<1)

Research Technicians Albert Gam – Biologist (2) Mona Febus – Microbiologist (3) Marc Alston – Biologist (2) Cherry Valerio – Biologist (2) Katia Dobrovolskaia – Visiting associate (2)

LIB staffing 1998-2003

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“Routine” regulatory activities Lot release Reference distribution Reference maintenance

semiannual checks replacement

Lot release activities 357 protocols submitted and

reviewed 1 withdrawn

Reference distribution 2002: 1978 vials in 107 shipments

sent to manufacturers

Lot release protocols submitted

Year Protocols

1999 4762000 4012001 4062002 357

Reference distribution

Year Shipments Vials

1999 104 19832000 146 24762001 101 21512002 107 1978


Operational issues

Operational issues Replacement of cat and ragweed

antisera

Transition to ISO compliance

Ragweed and cat antisera need to be replaced

S2a cat Released in 1998

S6 ragweed Released in 2000

Replacement programs for both initiated spring 2002

Immunization protocol

Allergen: Ragweed CatEwe #: 6747 7777

14-Jun-2002 Pre-bleed Plasmapheresis17-Jun-2002 Dose 1: 25 mcg Amb a 1 in CFA Dose 1: 25 mcg rFel d 1 in CFA

15-Jul-2002 Dose 2: 25 mcg Amb a 1 in IFA Dose 2: 25 mcg rFel d 1 in IFA

7-Aug-2002 Bleed Bleed12-Aug-2002 Dose 3: 25 mcg Amb a 1 in IFA Dose 3: 25 mcg rFel d 1 in IFA

4-Sep-2002 Bleed Bleed9-Sep-2002 Plasmapheresis Plasmapheresis

6/1 7/1 7/31 8/30 9/29

Immunization protocol

plasmapheresis

Immunization doses

6/1 7/1 7/31 8/30 9/29

Transmissible spongiform encephalopathies Animal

scrapie (sheep and goats) chronic wasting disease (mule deer,

elk) transmissible mink encephalopathy bovine spongiform encephalopathy

feline spongiform encephalopathy

Transmissible spongiform encephalopathies Human

kuru Creutzfeldt-Jakob disease

classic sporadic familial iatrogenic new variant (a/w bovine TSE)

Gerstmann-Sträussler-Scheincker fatal familial insomnia sporadic fatal insomnia

Scrapie In Europe for >250 years In US since 1947

>1000 flocks Vertical transmission Horizontal transmission, presumably by

contamination with placenta and blood during lambing season

Long incubation period No human transmission

Prion polymorphisms associate with different susceptibility, especially at codon 171 QQ (glutamine/glutamine) susceptible RR (arginine/arginine) resistant QR (glutamine/arginine) intermediate

Scrapie eradication program Preclinical testing and surveillance

Live animal test (third eyelid biopsy) May take months to years to turn positive

Tracking of infected and exposed animals

Cleanup strategies: identify and genotype exposed animals Destroy QQ exposed QR or RR exposed are tracked but may be

slaughtered for human consumption

Not believed to pose any risk to humans No recognized human transmission in three

centuries of exposure in Europe This serum is safe to use because

No documented transmission to humans Contact with affected sheep was limited

Not in adjoining pens; >30 feet distant Normal BSL 2 precautions in place for all work

with animal sera However, in order to maintain a serum

reagent that is as safe as possible…

Current approach… Begin immunizing two new sheep Process plasma from ewes 6747

and 7777 now Conserve current stocks

Current approach… If new sera are available before we

run out, sera from 6747 and 7777 will be saved frozen for possible future use

If we have a shortfall, sera from 6747 and/or 7777 will be used until the new sera are available

Current approach… Advantages

Highest degree of safety Large supplies for future

Disadvantages Possibility of two serum switches in

short period Time Expense

CBER Laboratory Quality Management Initiative

Seek ISO-17025 compliance for official product testing ISO – International Organization for

Standardization, Geneva, Switzerland 17025: “General requirements for the

competence of testing and calibration laboratories” (1999)Set of guidelines for labs that do testing and calibration to show operation of a quality system to assure technical competence and production of valid results

CBER Laboratory Quality Management Initiative

Establish policy with Center level Quality Manual

Audit for compliance with ISO-17025

Obtain Test/Lab Accreditation where appropriate “Accreditation – successful 3rd party audit

Why is CBER becoming ISO compliant? Establish recognized competence, assure

the trust and value of our data and processes

We require the manufacturers’ labs to be in GMP compliance International efforts at harmonization are

attempting to equate GMP and ISO requirements

ISO is an internationally recognized standard

Why is CBER becoming ISO compliant? (continued) Implement Laboratory Quality

Management policies and practices for official testing activities To document a high level of training,

competence, and proficiency To establish a consistent product

testing process

Elements of ISO compliance Management of

People Equipment Documents Processes

Elements of ISO compliance Management of People

Defined roles Appropriate training Demonstrated proficiency

Initial and ongoing Authorization process

Elements of ISO compliance Management of Equipment

Program for equipment calibration Maintenance Process to assure that only calibrated

and maintained equipment is used Assurance of measurement

traceability to accepted standards

Elements of ISO compliance Management of Documents

Includes policies, procedures, specifications, equipment manuals, certifications

Must be reviewed, issued and controlled

Approved and issued prior to use

Elements of ISO compliance Management of Processes

Approval of testing materials Environmental specifications and monitoring Handling of samples Validation and suitability Handling of data (including non-conforming

data) Corrective action, Preventive action systems Internal Audits Management review Recording and handling complaints

CBER’s commitment to implementation of a Lab Quality System

Establishment of CBER Quality Board Development of Quality Assurance

structure within CBER Appointment of Center Level Quality Manager Hiring Office quality Managers Appointment and hiring of Division Quality

Coordinators Preparation of CBER Quality Manual Purchase of Integrated Quality

Management computer software

What will this mean to LIB? No substantive protocol revisions Documentation

Formatting Substance

Formal separation of research and regulatory equipment

Internal audits External audits

Timetable (tentative) Implementation in stages over the

next 3 years… Seek accreditation in 2005 Software operational during 2003 Policies and Quality Manual issued 2003 Initiation of compliance audits (ongoing) Training and process development

(ongoing)


Research/regulatory update

Active research projects PI Slater

Cockroach allergen standardization Determination of optimal surrogate test Depletion analysis of CR extracts

Cockroach IgE combinatorial library Endotoxin in allergen vaccines

PI Rabin MDR proteins in T cell activation RSV responses in human tonsil

Publications

Patterson ML, Slater JE. Characterization and comparison of commercially available German and American cockroach allergen vaccines. Clin Exp Allergy 2002;32:721-727

Sutherland MF, Drew A, Rolland JM, Slater JE, Suphioglu C, O'Hehir RE. Specific monoclonal antibodies and human IgE show Hev b 5 is an abundant allergen in high protein powdered latex gloves. Clin Exp Allergy 2002;32:583-589

Trivedi B, Valerio C, Slater JE. Endotoxin content of standardized allergen vaccines. J Allergy Clin Immunol 2003 (in press).

Publications (reviews)

Lockey RF, Slater JE, Esch R. Preparation and standardization of allergen vaccines. In Middleton’s Allergy: Principles and Practice, 6th ed. St. Louis: Mosby (in press).

AbstractsRabin RL, Alston MA, Huang H, Slater JE. Cytokine secretion by

activated T cells is dependent on multidrug resistance protein-1 (MRP-1). J Allergy Clin Immunol 2003; 111:S153.

Valerio C, Slater JE. The effects of lipopolysaccharide (LPS) on

immune responses in C57Bl/6 mice. J Allergy Clin Immunol 2003; 111:S166.

Slater JE, Valerio C, Trivedi B. Endotoxin in standardized

allergen vaccines. J Allergy Clin Immunol 2003; 111:S243. Finlay WJJ, Rabin RL, Slater JE. Analysis of IgE heavy chain V-

gene usage in human tonsil. J Allergy Clin Immunol 2003; 111:S313.

Endotoxin content of allergen vaccines Allergenic extracts are not

required to undergo evaluation for the presence of pyrogens (21CFR 610.13(b))

Prior studies confirmed variable endotoxin content (Siraganian, et al. J Allergy Clin Immunol 1979; 64:526-533)

Products exempted in 21 CFR 610.13(b) Blood products Horse serum Bacterial, rickettsial and viral

vaccines Toxoids Toxins Allergenic extracts

LAL gel-clot method

Factor C Act. factor C

Factor B

Proclotting enzyme

Clotting enzyme

Factor GAct. factor B Act. factor G

Coagulogen Coagulin

ENDOTOXIN

–D- glucan

Endotoxin content of allergen vaccines – possible interference

Non-endotoxin (1,3)--D-glucans may induce clotting by an alternative pathway in the standard LAL assays

Proteases (especially in cat and mite extracts) may also induce clotting

Endotoxin content of allergen vaccines - approach Determine endotoxin content using

gel-clot method

Assess the contribution of non-endotoxin components ((1,3)--D-glucans, enzymes)

Study design Standardized allergen vaccines LAL gel-clot assay Adsorption of selected allergens with

ENP-silica resin, followed by LAL assay Endotoxin neutralizing protein (ENP) is a

~12 kDa, cationic, amphipathic protein that binds to and neutralizes the biological activity of lipopolysaccharide.

Pre-treat selected allergens at 95°C for 15 min, followed by LAL assay

Extract ManufacturerTotal gel-clot

activityDepleted gel-clot

activity

Corrected endotoxin content

(EU/mL)

Bermuda A 6--12 <6 8Bermuda E 12--24 Not tested 17Bermuda F 6--12 <6 8

June Grass A 12--24 <6 17June Grass B 60-120 6--12 76June Grass J 120-240 0 170

Meadow Fescue A 6--12 <6 8Meadow Fescue C 12--24 <6 17Meadow Fescue D 24-48 0 34Meadow Fescue F 12--24 0 17

Orchard A 60-120 <6 85Orchard C 12--24 <6 17Redtop E 24-48 <6 34Redtop G 24-48 <6 34

Ryegrass A 120-240 <6 170Ryegrass C 600-1200 320-600 410Ryegrass E 480-960 0 679Ryegrass F 12--24 Not tested 17Ryegrass G 1200-2400 96-192 1561

Sweet Vernal A 0.6-12 <6 3Sweet Vernal D 24-48 <6 26Sweet Vernal F 240-480 0 339

Timothy A 60-120 6--12 76Timothy D 6--60 6--12 10

GP

GP

GP

GP

GP

GP

GPGP

GP

GG

GG

G

G

G

G

G

G

GG

GG

P



activity


(EU/mL)

Ragweed Mix A 48-60 12--24 37Ragweed Mix B 60-120 32-60 41

Short Ragweed A 6--12 Not tested 8Short Ragweed B 60-120 32-60 41Short Ragweed C 600-1200 24-48 815Short Ragweed D 240-480 48-96 339Short Ragweed E 60-96 32-60 76Short Ragweed F 6--60 Not tested 19Short Ragweed J 1200-2400 0 1697

GP

G

G

G

GG

P

P

P



activity


(EU/mL)

Cat Pelt A 4800-6000 60-600 5177Cat Pelt A 24000-48000 96-192 33805Cat Pelt G 4800-6000 60-600 5177Cat Pelt G 4800-9600 6--12 6780Cat Hair B 240-480 0 339Cat Hair C 0.06-6 Not tested 1Cat Hair E 12000-24000 <6 16962Cat Hair F 600-1200 0 849Cat Hair H 240-480 6--60 339Cat Hair I 0.06-6 Not tested 1Cat Hair J 1200-2400 6--12 1689

GGG

GGG

G

GP

GP



activity


(EU/mL)

D.farinae A 6000-12000 60-600 8296D.farinae B 1200-2400 <6 1697D.farinae D 6000-12000 0 8485D.farinae E 4800-9600 0 6788D.farinae G 4800-6000 6--60 5367D.farinae I 600-1200 <6 849D.farinae J 600-1200 0 849

D.pteronyssinus A 6--12 <6 8D.pteronyssinus B 0.6-6 Not tested 2D.pteronyssinus C 0 Not tested 1D.pteronyssinus D 0.6-6 Not tested 2D.pteronyssinus G 6--60 Not tested 19D.pteronyssinus I 6--12 <6 8D.pteronyssinus J 24--48 Not tested 34

GGGGG

G

GGGGG

G

GP

GP

0%

20%

40%

60%

80%

100%

Gel

-clo

t act

ivity

from

(1,

3)-

-D

-glu

cans

Heat inactivationHeat inactivated

Extract Manufacturer EU/ml EU/ml

Ryegrass G 1200-2400 1200-2400Short Ragweed J 1200-2400 1200-2400Cat Pelt A 24000-48000 24000-48000Cat Pelt G 4800-9600 4800-9600D. farinae D 6000-12000 6000-12000D. farinae E 4800-9600 4800-9600D.pteronyssinus G 24-48 24-48D.pteronyssinus J 24-48 24-48

0

1

10

100

1000

10000

100000C

orr

ecte

d e

nd

oto

xin

co

nte

nt

(EU

/mL

)

0

1

10

100

1000

10000

100000C

orr

ecte

d e

nd

oto

xin

co

nte

nt

(EU

/mL

)

p = 0.02 p = 0.0003

Conclusions The observed LAL gel-clot activity

probably represents real endotoxin content, not -glucan or protease activity

Is this amount of endotoxin physiologically significant? Mean endotoxin content: 1,900

EU/mL (allergen immunotherapy dose 0.5 – 2.0 mL per month) 40 – 50 EU/kg (2,800 to 3,500 EU)

(administered IV) can elicit a rise in temperature, heart rate, and white blood cell count

Wolff SM. J Infect Dis 1973; 128:Suppl-64.

Michie HR, et al. N Engl J Med 1988 Jun 9 318;1481-6.

Specific endotoxin limits Product-specific Generally based on

Drug dose 5.0 EU/kg limit

Specific endotoxin limits

Product Limits (EU/dose)Vaccines 25 - 3000Penicilloyl polylysine 583*Doxorubicin HCl 285*Fentanyl citrate 80*Insulin (human) 56*

* Based on USP limits and estimated maximum therapeutic doses

The clinical consequences of endotoxin in allergen vaccines have not been studied

No data that adverse events from IT are associated with endotoxin levels

No data to support a beneficial effect of endotoxins

Future studies of allergen IT should be controlled for endotoxin dose

Role of endotoxin in safety and efficacy of IT should be assessed

Conclusions Pollens < cat and mite Cat hair < cat pelt D. pteronyssinus << D. farinae

Bioburden? Endogenous heat-stable ENP-binding

LAL activator in D. farinae? Endogenous ENP in D. pteronyssinus?

Plan Expand study of endotoxin content

Additional standardized and non-standardized extracts

Different methods (GC mass spec) Investigate differences between D.

farinae and D. pteronyssinus

Cockroach allergen standardizationClinical studiesDeveloping the appropriate surrogate

CorrelationDepletion studies

IgE combinatorial library to cockroach

Cockroach allergen standardization Clinical studies Developing the appropriate surrogate

Correlation Depletion studies

IgE combinatorial library to cockroach

Current standardized allergens D. farinae D. pteronyssinus Cat hair Cat pelt Short ragweed pollen Hymenoptera

Honey bee Wasp Yellow jacket Yellow hornet White-faced hornet Mixed vespid

Grass pollens Bermuda grass Red top June (Kentucky

blue) Perennial rye Orchard Timothy Meadow fescue Sweet vernal

Allergen standardization Establish a US standard, and Establish a testing procedure

Manufacturers may use the established procedure, or may develop equivalent procedures

Which allergens should be standardized? Impact criteria… Availability of stable, preferably lyophilized

material for use as long-term reference extracts. Consistency of currently marketed product. Widespread use as a diagnostic and/or

therapeutic reagent in the U.S. Number of manufacturers producing the product. Potential use in immunotherapy or diagnostics. Public health impact of correct diagnosis and/or

adequate treatment.

Allergens and asthma

Indoor allergens dust mites* cat* cockroach molds dog

Outdoor allergens molds * = already standardized

Cockroaches

Why is cockroach allergy important?

Ubiquitous Difficult to control Associated with asthma

Why is cockroach allergen standardization important? To the patient

More accurate diagnosis Safer and more effective immunotherapy

To the physician/scientist Better science (if you can’t measure it, you

can’t study it…) Pathophysiology Epidemiology Environmental control

To the FDA Safer, more effective product

Phase I - Laboratory

Develop/adapt methods for allergen determination

Compare allergen content of different lots

Goals Determine consistency of available

US products: protein content specific allergen content overall allergenicity

Determine best lot release measures

Extracts used as reference E2-Cg and E2-Ca Previously characterized Limited skin test data Lyophilized; available in large

quantities

Cockroach extracts studied

From all nine allergen extract manufacturers

aqueous 50% glyAmerican 4 11German 2 9

Relative Potency of All Cockroach Extracts (determined by competition

ELISA)

RP RP (scaled)

American Mean 0.07 0.05SD 0.05 0.04

Range 0 to 0.15 0 to 0.12

German Mean 0.05 0.22SD 0.06 0.19

Range 0 to 0.18 0.01 to 0.46

Aqueous Mean 0.00 0.03

SD 0.00 0.05Range

Gly

ce

rin

ate

d

Bla g 1 and Bla g 2 Levels in glycerinated German cockroach

Bla g 1 (U/mL) Bla g 2 (g/mL) Mean 3503 44 SD 1094 22 Range 2218 – 4854 8 - 66 E2 – Cg 13829 8

Correlation of protein concentrations and ELISA results

Protein concentration

Bla g 1 levels Bla g 2 levels

Relative potency

R2 = 0.83R

2 = 0

.92

R2 = 0.93

R2 =

0.9

6

R2 = 0.92

R 2 = 0.92

Conclusions Commercially available cockroach

allergen extracts: vary widely in protein content, Bla g 2

content, SDS-PAGE banding pattern, and overall allergenicity.

appear to be less potent and contain less Bla g 1 than the candidate reference extracts

Conclusions Established that cockroach

allergen vaccines need to be standardized

What we need now… New cockroach references Characterize the references ID50EAL testing (Intradermal Dilution for

50 mm Sum of Erythema Determines

Bioequivalent Allergy Units) Proceed to next phase

Phase II - Clinical

skin testing, histamine release, IT data

establish biological unitage and ideal dosing ranges

ID50EAL testing

Proficiency Recruitment Testing Analysis

ID50EAL testing

Recruitment Inclusion criteria

18 to 65 years of age history of allergic disease, such as

allergic rhinitis, related to exposure to the allergen of interest

puncture sum of erythema diameter responses (ΣE) to the allergen concentrate of ≥30 mm.

Recruitment Exclusion criteria

Asthma with use of systemic steroids in the past 12 months

Peak flow < 75% predicted at the time of testing Skin coloring or condition that would preclude the

measurement of erythema responses Dermographism (> 4 mm ΣE following saline skin test) Immunotherapy – past or present - with the test

allergen Current use of antihistamines, tricyclic

antidepressants, MAO inhibitors, and beta-blockers

How many study subjects?

22/11

2

2

zzn

n 1.0 0.05 26

0.20 181.5 0.05 59

0.20 392.0 0.05 104

0.20 69

Rabin et al., Sample Size Considerations for Establishing Clinical Bioequivalence of Allergen Formulations. Arb.Paul Ehrlich Inst.Bundesamt Sera Impfstoffe Frankf.A.M, in press

for D50 should be 10%

BAU/mL= 3-(14 - mean D50) * 100,000

D50 BAU/mL % change14 100000

12.6 21480 -79%15.4 465554 366%

10 12359 412 -67%

11 3704 200%8 137

7.2 57 -58%8.8 330 141%

for D50 is about 10-20%

Smith et al. Annals Allergy Asthma Immunol 1995; 75:317-323

for D50 is about 10-20%Using E no minimumGerman AmericanBAU E D50 BAU E D50

248892 82 14.83 8350 75 11.746928 74 11.57 440670 92 15.35

460467 107 15.39 111 77 7.811472 60 10.16 761 84 9.562723 36 10.72 1276 25 10.03980 47 9.79 218 31 8.42

1143 32 9.93 888 58 9.7213 60 8.4 13841 46 12.2

33333 67 13 1916 58 10.4639 46 9.4 88 61 7.6796 61 9.6

mean = 4429 mean = 11.16 mean = 1681 mean = 10.2895% LL 998 SD = 2.29 95% LL 342 SD = 2.3495% UL 19644 conf = 1.36 95% UL 8260 conf = 1.45

How many study subjects?

Estimate n = 40 to establish D50 for each extract (based on /= 1.5): 80

Geographic diversity: 80 x 3 = 240 Overlap between American CR and

German CR allergic subjects may permit reduction in n (150-200)

n 1.5 0.20 39



Need to establish the potency of candidate US reference materials by bioassay (ID50EAL)

NIAID Inner City Asthma Consortium

“…established in FY 2002 to explore and evaluate promising new strategies for the treatment of asthma among minority children residing in the inner city. This consortium of basic scientists and clinical investigators will conduct clinical studies to elucidate the immunopathogenesis and natural history of asthma in this population.”

From the FY 2003 Budget Justification Narrative, NIAID, http://www.niaid.nih.gov/director/congress/2002/cj/narrative.htm

NIAID Inner City Asthma Consortium steering committee

Busse, William W., MD Chair

Adams, Kenneth, PhD Eggleston, Peyton A., MD Gruchalla

, Rebecca S., MD, PhD Kattan, Meyer, MD Kercsmar, Carolyn M., MD Liu, Andrew H., MD Malveaux, Floyd J., MD, PhD Mitchell, Herman, PhD

Morgan, Wayne, MD, CM

O'Connor, George T., MD, MS

Pongracic, Jacqueline A., MD

Sampson, Hugh A., MD Smartt, Ernestine, RN Strunk, Robert C., MD Szefler, Stanley J., MD

Timetable Steering committee approval - done Study centers identified Order extracts IRB approvals IND approval Distribute materials Proficiency testing Proceed with study

Endotoxin content of cockroach vaccines

Manufacturer glycerol EU/mLAmerican cockroach D yes 60000-120000

E yes 9600-19200B yes 24000-48000A yes 600-1200F yes 12000-24000G yes 1200-2400J 60-120E 240-480C 6-12C 600-1200G 120-240

German cockroach F yes 12000-24000A yes 60000-120000C 240-480E 24000-48000G yes 600-1200C 48000-96000



Need to establish the potency of candidate US reference materials by bioassay (ID50EAL)

Endotoxin issue to be studied in depth

Will overall allergenicity measurements be sensitive to changes in specific allergen levels?• In mite stability study (1998-

1999), RP was stable at -20°C and 4°C for up to 12 months

Degradation of specific allergens (group 1 and 2; specific bands) was observed at 4°C

Soldatova LN, Paupore EJ, Burk SH, Pastor RW, Slater JE. J Allergy Clin Immunol 2000; 105:482-8.

RID with monospecific antiserum Examples: cat,

ragweed Advantages

quantitative monospecific

Disadvantages need to identify

relevant allergen(s)

Competition ELISA with pooled allergic human sera

Examples: mites, grasses Advantages

quantitative reflects spectrum of allergen

recognition does not require identification

of relevant allergens Disadvantages

use of pooled sera effects of fluctuations in

individual allergens difficult to measure

Specific loss of a single allergen

0

0.1

0.2

0.3

0.4

0.5

-6-4-20

log dilution

resp

onse

(a

bsor

banc

e)

Will overall allergenicity measurements be sensitive to changes in specific allergen levels?

Depletion analysis Raise specific antibodies to Bla g 1, 2,

4 and 5 Selectively adsorb Test for specific allergen levels Test for overall allergenicity

SDS-Page of Bla g 1 Absorbed Cockroach Antigen

Cr Sham Abs

Selective adsorption of Bla g 1

0

0.5

1

1.5

2

2.5

3

1 10 100 1000 10000

Dilution

Ab

sorb

ance

(45

0 n

m)

Bla g 1 ELISA

Sham adsorbedAdsorbed with anti-Bla g 1

Bla g 1 Bla g 2 Bla g 5Pre-immune 149.3 2.7 0.17anti-Bla g 1 14.8 4.1 0.07

Selective adsorption of Bla g 1 does not reduce the RP as measured by competition ELISA

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

1 10 100 1000 10000 100000

Dilution

Ab

sorb

ance

(45

0nm

)

Conclusions Established that cockroach allergen

vaccines need to be standardized Need to establish the potency of

candidate US reference materials by bioassay (ID50EAL)

Endotoxin issue to be studied in depth

Surrogate test may not be the competition ELISA

Conclusions Standardized German and American

cockroach allergen vaccines will facilitate definitive studies on the role

of cockroach allergens in inner city asthma, and on the best methods for eradication and treatment

make for safer and more effective products

laboratory of immunobiochemistry allergenic products advisory committee, april 8, 2003

Documents

marc alston biologist

cherry valerio biologist

replacement programs

md senior staff fellow

cat antisera

mona febus microbiologist

ronald rabin

ragweed antiseratransition