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Purification of a High Molecular Mass Protein in Streptococcus mutans

Purification of a High Molecular Mass Protein in Streptococcus mutans

Published on: September 14, 2019

Specific chemiluminescent protocol for dual-site recognition of Streptococcus mutans utilizing strong affinity

Xiaoxiao Su¹, Mengyao Wang¹, Yue Wu¹

¹Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.

|January 10, 2018

Abstract:

A novel dual-site recognition protocol was developed for chemiluminescent (CL) detection of Streptococcus mutans (S. mutans) based on a designed antibiotic-affinity strategy. Teicoplanin, a broad-spectrum antibiotic against Gram-positive bacteria, was adopted to functionalize magnetic particles and recognize S. mutans utilizing the strong affinity between this agent and D-Alanyl-D-Alanine peptide moieties in the bacterial cell wall. To achieve ideal specificity for S. mutans detection, rat immunoglobulin G2a (rat IgG2a) tagged with horseradish peroxidase (HRP) was used as the second recognition agent and signal tracer since Fab region of rat IgG2a could bind with streptococcal protein G highly expressed in the cell wall of S. mutans. Thus HRP-tagged sandwich complex of teicoplanin/S. mutans/rat IgG2a was formed on the magnetic particles, followed by a CL quantification of S. mutans based on a HRP-catalyzed luminol-H₂O₂-p-iodophenol CL reaction. This dual-site recognition protocol showed a linear range of 1.0 × 10²-1.0 × 10⁶ CFU mL^-1 and a detection limit of 33 CFU mL^-1 for S. mutans detection. The whole detection process could be completed within 70min. The recovery tests for food, environmental, pharmaceutical and biological samples showed acceptable recovery values between 83.0% and 110.0%, demonstrating its application potential for detection of bacteria in various sample matrixes.

Keywords:

Chemiluminescent Magnetic particles Rat immunoglobulin G2a Streptococcus mutans Teicoplanin

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Affinity and Avidity

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Titration Calculations: Strong Acid - Strong Base

Titration Calculations: Strong Acid - Strong Base

Calculating pH for Titration Solutions: Strong Acid/Strong Base
A titration is carried out for 25.00 mL of 0.100 M HCl (strong acid) with 0.100 M of a strong base NaOH. The pH at different volumes of added base solution can be calculated as follows:
(a) Titrant volume = 0 mL. The solution pH is due to the acid ionization of HCl. Because this is a strong acid, the ionization is complete and the hydronium ion molarity is 0.100 M. The pH of the solution is then:

Electron Affinity

Electron Affinity

The electron affinity (EA) is the energy change for adding an electron to a gaseous atom to form an anion (negative ion).

Strong Acid and Base Solutions

Strong Acid and Base Solutions

A strong acid is a compound that dissociates completely in an aqueous solution and produces a concentration of hydronium ions equal to the initial concentration of acid. For example, 0.20 M hydrobromic acid will dissociate completely in water and produces 0.20 M of hydronium ions and 0.20 M of bromide ions.

Titration of a Strong Acid with a Strong Base

Titration of a Strong Acid with a Strong Base

During the titration of a strong acid with a strong base, pH calculations are primarily based on the concentration of residual hydronium or hydroxide ions. Initially, a strong acid like hydrochloric acid fully dissociates, creating hydronium and chloride ions, resulting in a low pH. The addition of a strong base like sodium hydroxide alters the concentration of hydronium ions by neutralizing them. As more base is added, the pH gradually increases. At the equivalence point, all hydronium ions...

Titration Calculations: Weak Acid - Strong Base

Titration Calculations: Weak Acid - Strong Base

Calculating pH for Titration Solutions: Weak Acid/Strong Base
For the titration of 25.00 mL of 0.100 M CH3CO2H with 0.100 M NaOH, the reaction can be represented as:

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