Author details

4. Discussion

process control have no be reported.

Innovations in Cell Research and Therapy

fractions, gammaglobulin) [29].

maceutical industry [30, 31].

5. Conclusions

RIA methods (5 h).

or RIA (\$ 25/test).

62

and reliability for this purpose.

The availability of a quantitation method for human plasma, Cohn fractions, and

A rapid and low-cost flow cytometry test applied in the control of the industrial

According to this, the threshold of the assay was adjusted to select the level of antibody content, which will be accepted, to produce an immunoglobulin with a potency of 250 μg/vial (1250 IU/vial), according to the regulatory requirements [13, 14, 28]. To validate the assay, chemical and physical-chemical factors that affect the binding of antigen-antibody influence in the red blood cells were determined. The spatial conditions of cell packing did not affect the assay. The adsorption of antibodies to the cell surface was 1000 times greater when the incubation temperature was 37°C without this affecting the dissociation rate [9]. The different fractions behave similarly in the assay conditions, and there was no interference by autoagglutination at the concentration of red blood cells used (Figure 7). The analytical quality assurance of the trial showed that the method presents good reproducibility, repeatability (1–7.5 VC%), and correlation with the standard curve (r: 0.9267). The assay is specific and recovery is 95% of the known Anti-D concentration values. For validation our strategy was based on an international guide accepted by manufacturers and control laboratories [13, 30]. The validation method meets the requirements according to the good manufacturing practices in the pharmaceutical industry. A validation methodology was followed to meet the special requirements of standardization of the good manufacturing practices in the phar-

Biological variability, red blood cells, and matrices were considered in a valida-

This flow cytometry method is a sensitive and specific method that allows reproducible results. The concentration values are comparable with those estimated with other methods such as RIA and EIA used by commercial manufacturers of gamma globulins, and the method presents accuracy and precision. The test is completed in 3 h and is easy to perform, allowing quantitative assessment of Anti-D antibodies from plasma, fractions of the Cohn process, and finished products. The flow cytometry method presented shorter processing times (3 h) than the EIA or

The potential of the flow cytometry method described here represents an alter-

The cost of materials is slightly lower for flow cytometry (\$ 15/test) than in EIA

native to quantify Anti-D in different matrices and meets the criteria of good laboratory practices (simple, fast, and reliable as well as being sensitive and accurate). In the control of industrial processes, this method has shown reproducibility

tion protocol according to international guidelines [14, 30].

Anti-D gammaglobulin has a relevant importance for the process control of the Anti-D IgG production. Pharmaceutical companies are employing sensitive EIA, RIA, and flow cytometry techniques [10, 11, 15, 20, 24] for the quantitation of Anti-D level in finished products, the use in Anti-D gammaglobulin manufacturing

process of large-scale production obtaining Anti-D gammaglobulin has been described. This responded to the need of an alternative method to EIA and RIA in the quantitation of Anti-D in different process samples (human plasma, Chon

Sergio A. Oviedo1,2

1 Blood Products Laboratory National University of Cordoba, Córdoba, Argentina

2 Group Quality and Regulatory Affairs, School of Chemistry, Catholic University of Cordoba, Argentina

\*Address all correspondence to: sergio.oviedo@unc.edu.ar

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
