BIOPHEN PROTEIN C5, C2.5, MODELS 221205, 221202

K050365 · Hyphen Biomed · GGP · Dec 27, 2005 · Hematology

Device Facts

• Record ID: K050365
• Device Name: BIOPHEN PROTEIN C5, C2.5, MODELS 221205, 221202
• Applicant: Hyphen Biomed
• Product Code: GGP · Hematology
• Decision Date: Dec 27, 2005
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7290
• Device Class: Class 2

Indications for Use

Biophen Protein C 5 & 2.5 kit is a Chromogenic assay for measuring the Protein C Activity in human citrated plasma using a manual or an automated method.

Device Story

Biophen Protein C is an in-vitro diagnostic kit for measuring Protein C activity in human citrated plasma. The assay utilizes Protac, an enzyme extracted from Agkistrodon C Contortrix snake venom, to activate Protein C in the sample. The activated Protein C (APC) then cleaves a specific chromogenic substrate (SaPC-21), releasing para-nitroaniline (pNA). The resulting color change is measured spectrophotometrically at 405nm. The intensity of the color is directly proportional to the Protein C activity in the plasma. The device is intended for use in clinical laboratory settings by trained personnel. Results assist clinicians in assessing Protein C levels, which are essential for regulating coagulation. The kit is provided in lyophilized form and requires reconstitution before use.

Clinical Evidence

Bench testing only. Precision studies showed intra-assay CVs of 0.37-1.17% and inter-assay CVs of 1.26-1.97%. Linearity was confirmed with r² values of 0.996 and 0.999 across a 0-100% activity range. Limit of detection is 5% activity. Method comparison studies against the predicate device at two sites (n=21 and n=17) yielded r²=0.99, confirming analytical equivalence.

Technological Characteristics

Chromogenic assay. Reagents: Protac (Agkistrodon C Contortrix venom enzyme) and SaPC-21 chromogenic substrate. Form: Lyophilized. Measurement: Spectrophotometric at 405nm. Methodology: Manual or automated. Stability: Reconstituted reagents stable 3 days at room temperature or 3 months at 2-8°C.

Indications for Use

Indicated for the quantitative determination of Protein C activity in human citrated plasma using manual or automated methods.

Predicate Devices

• Coamatic Protein C (K922201)

Submission Summary (Full Text)

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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
K050365

B. Purpose for Submission:
New Device

C. Measurand:
Protein C

D. Type of Test:
Quantitative

E. Applicant:
Hyphen BioMed

F. Proprietary and Established Names:
Biophen Protein C

G. Regulatory Information:
1. Regulation section:
21 CFR 864.7290, Factor Deficiency Test
2. Classification:
Class II
3. Product code:
GGP, Test, Qualitative and Quantitative Factor Deficiency
4. Panel:
(81) Hematology

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H. Intended Use:

1. Intended use(s):

Biophen Protein C (5 & 2.5) kit is an in-vitro diagnostic test for the quantitative determination of Protein C in human citrated plasma by chromogenic assay, using a manual or automated method.

2. Indication(s) for use:

Biophen Protein C is a chromogenic assay for measuring the Protein C activity in human citrated plasma using a manual or automated method.

3. Special conditions for use statement(s):

Not applicable.

4. Special instrument requirements:

Not applicable.

I. Device Description:

Biophen Protein C in-vitro diagnostic kit is a chromogenic assay for measuring the Protein C activity in human plasma using a manual or automated method. The kit contains a chromogenic substrate and Protein C activator. The kit is marketed in two sizes, Biophen PC, 5 [4 vials reconstituted to 5ml (200 tests)] and Biophen PC, 2.5 [3 vials reconstituted to 2.5ml (75 tests)].

J. Substantial Equivalence Information:

1. Predicate device name(s):

Coamatic Protein C

2. Predicate 510(k) number(s):

K922201

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3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| | Biophen Protein C | Coamatic® Protein C |
| Intended use | Used for measuring the Protein C activity in human plasma by chromogenic assay using a manual or an automated method. | Same |
| Form | Lyophilized | Same |
| Matrix | Reagent 1: Protac® in distilled water.
Reagent 2: SaPC-21 substrate in distilled water. | Reagent 1: Protein C activator in distilled water.
Reagent 2: S-2366 Substrate in distilled water. |
| Materials | Reagent 1 (Protac®): purified enzyme extracted from the Agkistrodon C Contortrix snake venom, lyophilized.
Reagent 2 (SaPC-21): Chromogenic substrate specific for Protein C, lyophilized. | Reagent 1 (Protein C Activator): Lyophilized venom enzyme from southern copperhead snake (Agkistrodon C. Contortrix).
Reagent 2 (S-2366): Chromogenic substrate, lyophilized. |

K. Standard/Guidance Document Referenced (if applicable):

Not provided.

L. Test Principle:

Protein C is an anticoagulant protein, which is dependant on Vitamin K. It inhibits and regulates coagulation through the specific cleavage of factors Va and VIIIa, suppressing their procoagulant cofactor activity.

Protein C in human plasma is measured following specific activation using Protac®, an enzyme extracted from snake venom (Agkistrodon C Contortrix). The activated protein C (APC) then specifically cleaves the specific substrate SaPC-21, releasing

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para-nitroaniline (pNA). The release of para-nitroaniline produces color, which is measured at 405nm. There is direct relationship between release of par-nitroaniline and the protein C activity in the tested plasma.

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

A reproducibility study was performed on Protein C concentrations of 98%, 59% and 39%. The results are as follows:

| Mean Activity | Intra-assay (CV %) | Mean Activity | Inter-Assay (CV %) |
| --- | --- | --- | --- |
| 98% | 0.37% (n=9) | 98% | 1.26% (n=12) |
| 59% | 1.17% (n=10) | 59% | 1.97% (n=12) |
| 39% | 0.84% (n=10) | 30% | 1.51% (n=12) |

#### b. Linearity/assay reportable range:

Linearity was determined using the Protein C calibrator with values of 0, 20, 40, 60, and 100% activity. The linear regression results for two lots of Biophen Protein C reagent are as follows:

$$
\begin{array}{l}
r^{2} = 0.996 \quad y = 0.009x + 0.022 \\
r^{2} = 0.999 \quad y = 0.081x + 0.056 \\
\end{array}
$$

The highest reportable value is 100%.

#### c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Stability studies were performed on Protac (R1) and SaPC-21 substrate (R2) vials stored at room temperature (18-25°C) or at 2-8°C, reconstituted. The activity was tested, after storage periods of 1-4 weeks. In addition, they were tested at 3 and 7 months. This study determined that reconstituted R1 and R2 reagents are stable: 3 days at room temperature (18-25°C), 3 months at 2-8°C.

#### d. Detection limit:

Threshold detection testing was performed using two lots of reagent. The limit of detection was determined to be 5% activity.

#### e. Analytical specificity:

Not applicable.

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f. Assay cut-off:

Not applicable.

2. Comparison studies:

a. Method comparison with predicate device:

Comparison studies using the Biophen Protein C assay and the predicate device were performed at two sites with patient samples presented for Protein C testing. The results are as follows:

Site 1 $r^2 = 0.99$ y = 1.00x + 0.84 n = 21

Site 2 $r^2 = 0.99$ y = 0.95x + 7.9 n = 17

b. Matrix comparison:

Not applicable.

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

b. Clinical specificity:

Not applicable.

c. Other clinical supportive data (when a. and b. are not applicable):

Not applicable.

4. Clinical cut-off:

Not applicable.

5. Expected values/Reference range:

The expected range was established in the literature. The 100% Protein C concentration corresponds to the concentration in a normal human citrate plasma pool, obtained by pooling plasma from healthy males or females aged from 18 to 55 years, and out of any medication. The Protein C concentration in adults is usually between 70 and 140%.

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N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

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