DESIGN OUTPUT

HIV Ag/Ab ELISA

C

1 Product Design .............................................................................................................................................................. 2

1.1 Area of Applicability ................................................................................................................................................................... 2
1.2 Test Format .................................................................................................................................................................................... 2
1.3 Physical and Chemical Properties .......................................................................................................................................... 2
1.4 Classification and Dangerous Goods Aspects................................................................................................................... 5
1.5 Storage ............................................................................................................................................................................................ 5
1.6 Stability ........................................................................................................................................................................................... 5
2 Product and Quality Specifications ............................................................................................................................. 5
2.1 Description of Test Methods ................................................................................................................................................... 5
2.2 QC Specifications / COA ........................................................................................................................................................... 5

Form 4.2-01.06-02 1/7

Rev. 003 | valid of 07.04.2014
1 Product Design

1.1 Area of Applicability

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HUMAN HIV Ag/Ab ELISA has been designed as a double antigen and double antibody sandwich ELISA of the 4
generation for the detection of antibodies to HIV-1 (including group O) and HIV-2 and of the HIV-1 p24 antigen in
serum and plasma. Results are obtained by comparison with a cut-off value.

1.2 Test Format

HUMAN HIV Ag/Ab ELISA consists of 13 components, including coated microtiter strips [MIC] , negative and two
positive controls [NC], [PC-1], [PC-2], conjugates CON-1 , CON-2 , conjugate diluents [DIL-C1], [DIL-C2] wash solution
concentrate [WS], substrate reagent A (TMB) [SA], substrate reagent B (buffer) [SB], stop solution [STOP] and adhesive
strips. Test formats correspond with the respective customer needs. For OEM customers the reagents may also be
delivered in neutral unlabeled vials.

Cat. No.
Component Quantity / Volume
51215 512155
[MIC] Microtiter strips 1x 5x 12 x 8-well strips, sealed in aluminium- foil bag,
labelled
[NC] anti-HIV 1/2, p24 1 x 2.5 ml 2 x 2.5 ml filled into round glass vial, closed with green
Negative Control screw cap, labelled
[PC-1] anti-HIV 1/2 Positive 1 x 2.0 ml 2 x 2.0 ml filled into round glass vial, closed with red screw
Control cap, labelled
[PC-2] p24 Positive Control 1 x 2.0 ml 2 x 2.0 ml filled into round glass vial, closed with white
screw cap, labelled
[CON-1] anti-HIV 1/2, p24 1 x 1.2 ml 1 x 3.6 ml filled into round brown glass vial, closed with
Conjugate yellow screw cap, labelled
[CON-2] HRP Conjugate 1 x 2.0 ml 2 x 2.0 ml filled into round brown glass vial, closed with blue
screw cap, labelled
[DIL-C1] Conjugate-1 Diluent 1 x 12 ml 3 x 12 ml filled into round transparent plastic vial, closed
with transparent screw cap, labelled
[DIL-C2] Conjugate-2 Diluent 1 x 20 ml 2 x 20 ml filled into round transparent plastic vial, closed
with transparent screw cap, labelled
[WS] Wash solution 1 x 50 ml 2 x 120 ml concentrate, filled into round transparent plastic
vial, closed with transparent screw cap, labelled
[SA] Substrate Reagent A 1 x 2.5 ml 3 x 2.5 ml concentrate, filled into round brown plastic vial,
closed with brown screw cap, labelled
[SB] Substrate Reagent B 1 x 25 ml 3 x 25 ml filled into round brown plastic vial, closed with
brown screw cap, labelled
[STOP] Stop Solution 1 x 25 ml 2 x 50 ml filled into round transparent plastic vial, closed
with transparent screw cap, labelled
Adhesive strips 2 10

1.3 Physical and Chemical Properties

1.3.1 Microtiter strips [MIC]
Aggregate state solid, coated strips, breakable into single wells
Density n/a
Appearance clear and colourless polystyrene strips
Smell none
Chemical constituents recombinant antigens (E. coli; HIV-1: gp160, p31, gp41 HIV-1 group O,
HIV-2: gp36) and mouse monoclonal anti-p24 HIV-1 antibody

Design Output for HIV Ag/Ab ELISA 2/7

Rev. 003
1.3.2 Negative Control [NC]
Aggregate state liquid
Density n/a
Appearance green dyed, transparent or slightly opalescent
Smell none
Chemical constituents human plasma based negative control (heat inactivated, non-reactive for p24
HIV-1 antigen, anti-HIV-1/2, HBsAg and anti-HCV);
dye: 0.015% naphtol green; preservative: 0.01% thimerosal

1.3.3 Positive Control 1 [PC-1]

Aggregate state liquid
Density n/a
Appearance orange dyed, transparent or slightly opalescent
Smell none
Chemical constituents human plasma based control positive for anti-HIV-1/2 (heat inactivated, non-
reactive for p24 HIV-1 antigen, HBsAg and anti-HCV);
dyes: 0.005% amaranth, 0.01% naphtol yellow;
preservatives: 0.025% thimerosal; 0.01% gentamicin sulphate

1.3.4 Positive Control 2 [PC-2]

Aggregate state liquid
Density n/a
Appearance crimson-red dyed, transparent or slightly opalescent
Smell none
Chemical constituents purified recombinant antigen p24 HIV-1 in heat inactivated human plasma
(non-reactive for anti-HIV-1/2, HBsAg and anti-HCV, p24 HIV-1 antigen); dye:
0.05% phenol red; preservatives: 0.05% thimerosal; 0.01% gentamicin sulphate

1.3.5 Conjugate 1 (concentrate) [CON-1]

Aggregate state liquid
Density n/a
Appearance colourless to pale yellow, transparent or slightly opalescent
Smell none
Chemical constituents biotinylated recombinant antigens (E. coli; HIV-1: gp41, p31, gp41 HIV-1 group
O, HIV-2: gp36) and mouse monoclonal anti-p24 HIV-1;
buffer: tris 6.06 g/l, 0.8 ml/l HCl
preservatives: 0.01% thimerosal, 0.01% gentamicin sulphate

1.3.6 Conjugate 2 (concentrate) [CON-2]

Aggregate state liquid
Density n/a
Appearance colourless to pale yellow, transparent or slightly opalescent
Smell none
Chemical constituents horseradish peroxidase labelled HIV-1: gp41, HIV-2: gp36 and horseradish
peroxidase labelled streptavidin
glycerin 50%, Na2HPO4x12H2O, NaCl
preservative: 0.02% phenol

Design Output for HIV Ag/Ab ELISA 3/7

Rev. 003
1.3.7 Conjugate 1 Diluent [DIL-C1]
Aggregate state liquid
Density n/a
Appearance orange dyed, slightly opalescent; pellet, dissolved at shaking, may form
Smell none
Chemical constituents 29,25g/l NaCl, 0.5% bovine serum;
preservatives: 0.01% thimerosal, 0.01% gentamicin sulphate, 0.02% phenol

1.3.8 Conjugate 2 Diluent [DIL-C2]

Aggregate state liquid
Density similar to water
Appearance blue dyed, slightly opalescent
Smell none
Chemical constituents NaCl, urea, sodium caseinate
preservative: 0.01% thimerosal

1.3.9 Wash solution (25x concentrate) [WS]

Aggregate state liquid
Density n/a
Appearance colourless to pale yellow, slightly opalescent
Smell none
Chemical constituents 21.9 g/l Na2HPO4x12H2O, 295.0 g/l NaCl, 2.5% tween-20
preservatives: none

1.3.10Substrate reagent A [SA] (TMB)

Aggregate state liquid
Density n/a
Appearance colourless, transparent
Smell yes
Chemical constituents 4.0 g/l 3,3', 5,5'-tetramethylbenzidine (TMB); 70 % DMSO; preservatives: none

1.3.11Substrate reagent B [SB] (buffer)

Aggregate state liquid
Density similar to water
Appearance colourless, transparent
Smell none
pH 4.2 ± 0.1
Chemical constituents 0.03 mol/l citrate buffer, 0.008% hydrogen peroxide
Preservative: 0.04% ProClin 300

1.3.12Stop solution [STOP]

Aggregate state liquid
Density similar to water
Appearance colourless, transparent
Smell none
Chemical constituents 0.2 mol/l sulphuric acid

Design Output for HIV Ag/Ab ELISA 4/7

Rev. 003
1.4 Classification and Dangerous Goods Aspects
The stop solution contains 0.2 mol/l sulphuric acid (1.1% H2SO4). Being below the threshold of 5% no special
dangerous goods restrictions have to be observed. All the other reagents listed above are not to be treated as
dangerous goods.

1.5 Storage
All kit components have to be stored at 2...8°C. Freezing must be avoided.

1.6 Stability
HIV Ag/Ab ELISA is stable up to 24 months from the date of production when sealed and stored properly.

2 Product and Quality Specifications

2.1 Description of Test Methods

2.1.1 General remarks
Tests are performed by the quality control department according to written procedures. Testing equipment is
checked in advance and calibrated at suitable intervals.

2.1.2 Washing step

The plates are washed between the incubation steps with an automatic washer, programmed for this method. The
washer has been qualified in advance and is subject to routine maintenance.

2.1.3 Photometric measurement

ELISA strip/plate readers with wavelength of 450 nm and a reference wavelength of 620 nm are employed. For
other photometric measurements typical UV/VIS photometers are employed. Reproducibility and accuracy of
wavelengths are controlled according to the recommendations of the manufacturer.

2.1.4 Function test

The function test is performed according to the instruction for use.

2.2 QC Specifications / COA

QC specifications for product release are summarised in the attached Certificate of Analysis.

Design Output for HIV Ag/Ab ELISA 5/7

Rev. 003
 CERTIFICATE OF ANALYSIS

HIV Ag/Ab ELISA

ELISA for the detection of antibodies to
Human Immunodeficiency Virus Types 1
and 2 (HIV-1, HIV-2), HIV-1 group O and
p24 HIV-1 antigen in human serum and
plasma
Cat. No.: 51215
Pack. Lot: LOTNUMMER
Prod./Inv. Date: PRODDATUM
Expiry Date: EXPDATUM

Storage conditions: Store at 2...8°C !

Performance Test
Specifications Results

A Substrate blank Meet specifications passed

B MNC (mean negative control) Meet specifications passed
C MPC (mean positive control) Meet specifications passed
D QC patient panel In agreement with passed
predetermined status

MPC:MNC Within range passed

COV calculation Within range passed
Positive result Within range passed
Negative result Within range passed

This product met the quality control release criteria and has been released by the responsible
Quality Control Department of HUMAN GmbH.

Following release of the product, this certificate of analysis is electronically regenerated and
valid without signature.

Date of print: 23.3.2015

Design Output for HIV Ag/Ab ELISA 6/7

Rev. 003
 CERTIFICATE OF ANALYSIS

HIV Ag/Ab ELISA

ELISA for the detection of antibodies to
Human Immunodeficiency Virus Types 1
and 2 (HIV-1, HIV-2), HIV-1 group O and
p24 HIV-1 antigen in human serum and
plasma
Cat. No.: 512155
Pack. Lot: LOTNUMMER
Prod./Inv. Date: PRODDATUM
Expiry Date: EXPDATUM

Storage conditions: Store at 2...8°C !

Performance Test
Specifications Results

A Substrate blank Meet specifications passed

B MNC (mean negative control) Meet specifications passed
C MPC (mean positive control) Meet specifications passed
D QC patient panel In agreement with passed
predetermined status

MPC:MNC Within range passed

COV calculation Within range passed
Positive result Within range passed
Negative result Within range passed

This product met the quality control release criteria and has been released by the responsible
Quality Control Department of HUMAN GmbH.

Following release of the product, this certificate of analysis is electronically regenerated and
valid without signature.

Date of print: 23.3.2015

Design Output for HIV Ag/Ab ELISA 7/7

Rev. 003