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2016, Number 3

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Correo Científico Médico 2016; 20 (3)

Pertussis toxoid purification of Bordetella pertussis

Hernández MM, Rivas RED, Llanos RHD, Álvarez BMA

Language: Spanish
References: 11
Page: 468-478
PDF size: 383.51 Kb.


ABSTRACT

Introduction: pertussis toxoid is a widely used in acellular vaccines against pertussis protein. Develop a protocol for purification with few steps and with parameters suitable purification is very important for chemical characterization and assessment of adverse effects that can lead to inoculate mainly mice. Purification protocols described in scientific literature involve several purification steps and low yields.
Objective: to develop a protocol for purification of pertussis toxoid to ensure a high degree of purity and a yield of over 80% from cultures of a mutant of B. pertussis obtained at the National Center for Scientific Research (CNIC).
Methods: for purifying the toxoid cation a Fractogel EMD SO-3 cationic interchange column was used by increasing the pH and salt concentration to obtain the expected results. The total protein determination was performed using linearized macromethod of Bradfor10 using as bovine serum albumin standard (BSA).
Results: 3.28 mg toxoid was obtained per liter of culture with a yield higher than 86.7% and a purity of 96.7%.
Conclusions: the purification process described allows for the pertussis toxoid with a higher yield than 80% and high purity, ensuring the quality required for biochemical and immunological characterization.


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