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Related Concept Videos

Ion Exchange01:17

Ion Exchange

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Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or...
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Anionic Chain-Growth Polymerization: Overview01:20

Anionic Chain-Growth Polymerization: Overview

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The polymerization process that involves carbanion as an intermediate is called anionic polymerization. It is also a type of addition or chain-growth polymerization. Anionic polymerization gets initiated by a strong nucleophile such as an organolithium or a Grignard reagent. The most commonly used initiator for anionic polymerization is butyl lithium. Monomers involved in anionic polymerization must possess a vinyl group bonded to one or two electron-withdrawing groups. For instance,...
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Colloidal precipitates01:09

Colloidal precipitates

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The high insolubility of some precipitates can result in an unfavorable relative supersaturation. This can lead to colloidal particles with a large surface-to-mass ratio, where adsorption is promoted. For instance, in the precipitation of silver chloride, silver ions are adsorbed on the surface of the colloidal particles, forming a primary layer. This layer attracts ions of opposite charge (such as nitrate ions), forming a diffuse secondary layer of adsorbed ions. This electric double layer...
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Precipitation Processes01:12

Precipitation Processes

498
The experimental conditions in a gravimetric analysis should be optimized to maximize the particle size and purity of the obtained precipitate. Ideally, the concentration of the precipitating reagent should be low with effective stirring to maintain low relative supersaturation for the growth of large crystals. In homogeneous precipitation, the precipitant is slowly generated by a chemical reaction in the solution to avoid local reagent excesses. For example, urea decomposes gradually to...
498
Cationic Chain-Growth Polymerization: Mechanism00:57

Cationic Chain-Growth Polymerization: Mechanism

2.4K
The cationic polymerization mechanism consists of three steps: initiation, propagation, and termination. In the initiation step of the polymerization process, the π bond of a monomer gets protonated by the Lewis acid catalyst, which is formed from boron trifluoride and water. The protonation of the π bond generates a carbocation stabilized by the electron‐donating group. In the propagation step, the π bond of the second monomer acts as a nucleophile and attacks the...
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Washing, Drying, and Ignition of Precipitates00:52

Washing, Drying, and Ignition of Precipitates

988
After filtration, the precipitate is washed to remove coprecipitated impurities and any remaining mother liquor. Colloidal precipitates, such as silver chloride, are washed with an electrolyte (such as dilute nitric acid) to prevent the peptization of the precipitate. In the case of slightly soluble precipitates, the wash solution contains a common ion to reduce solubility. Lead sulfate, which is slightly soluble in water, is washed with dilute sulfuric acid. Similarly, wash solutions may be...
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Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides
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Cationic polymer precipitation for enhanced impurity removal in downstream processing.

Zhao Li1, Justin Chen1, Kirby Martinez-Fonts2

  • 1Biologics Process Development, Biologics Process Research and Development, Merck & Co., Inc., Rahway, New Jersey, USA.

Biotechnology and Bioengineering
|May 6, 2023
PubMed
Summary
This summary is machine-generated.

Polyallylamine (PAA) precipitation effectively removes host cell proteins (HCPs) and DNA from biologics, enhancing polysorbate stability and extending shelf life with minimal product loss.

Keywords:
PS-80 stabilitydownstream purificationlipase removalmass spectrometryprecipitationshelf life

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Downstream Process Development
  • Protein Purification

Background:

  • Impurities like host cell proteins (HCPs) and DNA can compromise biologic product quality and stability.
  • Early-stage impurity removal is crucial for efficient downstream purification processes.
  • Polysorbate excipient stability is vital for maintaining drug efficacy and shelf life.

Purpose of the Study:

  • To evaluate polyallylamine (PAA) precipitation for enhanced host cell protein (HCP) removal in biologic purification.
  • To assess the impact of PAA precipitation on polysorbate excipient stability and product shelf life.
  • To optimize PAA precipitation conditions using high-throughput screening and process analytical tools (PATs).

Main Methods:

  • High-throughput screening of PAA precipitation conditions (pH, conductivity, concentration).
  • Utilized process analytical tools (PATs) for particle size analysis.
  • Scaled up precipitation to 20L and characterized impurity removal using ELISA, mass spectrometry, and DNA assays.
  • Analyzed residual PAA and product quality attributes.

Main Results:

  • Achieved >75% reduction in HCP concentration, >90% reduction in HCP species, and >99.8% reduction in DNA.
  • Improved polysorbate-containing formulation buffer stability by at least 25% for three monoclonal antibodies (mAbs).
  • Observed minimal impact on product quality with <5% yield loss and residual PAA <9 ppm.

Conclusions:

  • PAA precipitation is a valuable tool for enhancing HCP and DNA clearance in biologic downstream purification.
  • This method improves excipient stability, enabling longer product shelf life.
  • PAA precipitation integrates effectively with existing purification platforms, offering solutions for challenging purification programs.