**Acknowledgements**

I am very thankful to my all students; Ahlem Lamnai, Souad M'hamedi, Saadia Benmadi, Wasila Aeid.

## **Disclosures**

*Biochemical Testing - Clinical correlation and Diagnosis*

the holes can be distributed evenly.

*End point assessment* Optical density by

6.Pasteur pipette as well cutters to produce holes approximately 5 mm diameter. If single holes are to be punched, a grid should be placed under the gel so that

Since Thomas A. E. Platts-Mills and Kimishige Ishizaka have discovered that fresh normal human serum in EGTA buffer was found to cause >90% hemolysis of unsensitized rabbit red blood cells (RaRBC) [11], simple timed lysis assay and dilution methods called AP50 (Alternative Pathway 50) was performed to quantify hemolytic complement activity in human serum [13–15]. This reaction requires C3, factors B and D, and Mg++ ions to form the C3 convertase (C3bBb) [14, 15]. In AP50, ACP is activated and measured by virtue of RaRBC decreased sialic acid content in addition to the blockade of the classical pathway activation by chelation of calcium by EGTA [16]. Nonetheless, AH50 requires a lot of material like spectrophotometer and consumable test tubes and highly skillful personal. To adapt this assay to simple labs, a semi-solid phase assay was proposed to measure ACP activity by using chicken erythrocytes incorporated in agarose gel called AP100. Cell membranes of chicken erythrocytes have the same properties as those of the

*Number of samples* <5 per day Several samples series per day *ACP Activators* Red cells Red cells + agarose particles

*Transport* No Yes (respecting the cold chain)

*Others* • More suitable complement related

Spectrophotometer

*Laboratory work flow steps* +++ Less *Time consuming* +++ Less *Storage* No 15–21 days

*Global cost* +++ Less *Assay related error* +++ Less *Manipulator linked error* +++ Less

*Hemolytic agarose assay features in comparison to tube hemolytic assay.*

**Fluid phase assay (AH50) Semi-solid phase assay (AP100)**

Diameter measurement directly

diseases screening

ogy handling

• Do not need a deep immunotechnol-

• Storage of utilized plates for *intra* and *inter*-laboratory comparisons

4.Level table.

5. 37°C incubator.

7.Refrigerated centrifuge.

9.pH meter/temperature probe.

8.Electronic balance.

**4. Discussion**

**108**

**Table 2.**

The author has nothing to disclose.
