Optimized recovery of monoclonal antibodies from transgenic goat milk by microfiltration
- 20 July 2004
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 87 (3), 274-285
- https://doi.org/10.1002/bit.20112
Abstract
The Predictive Aggregate Transport Model for microfiltration is used in combination with optimum fluid mechanics and electrostatics to maximize recovery of a heterologous immunoglobulin (IgG) from transgenic goat milk. The optimization algorithm involved varying pH (6.8–9), transmembrane pressure (2–4.5 psi), milk feed concentration (1–2X), membrane module type (linear vs. helical design), and axial velocity (Reynolds number: 830‐1170). Operation in the pressure‐dependent regime at low uniform transmembrane pressures (≈2 psi) using permeate circulation in co‐flow, at the pI of the protein (9 in this case) was used to increase IgG recovery from less than 1% to over 95%. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and attenuated total reflection Fourier transform infrared spectroscopy of the microfiltration permeate samples confirmed that all the fat globules and most of the casein micelles were retained in the MF membrane whereas a large amount of the target IgG was transported through the membrane. Transmembrane pressure and hence permeation flux was kept low (≈15 lmh) to maximize IgG membrane transport and thus recovery, due to a sparse deposit on the membrane which facilitated high solute transport. Next, an analytical method was used to optimize the diafiltration process using the aggregate transport model, experimental target protein sieving coefficients and permeation flux (Baruah and Belfort, 2003). The methodology reported here should be generalizable to the recovery of target proteins found in other complex suspensions of biological origin using the microfiltration process.Keywords
This publication has 16 references indexed in Scilit:
- A Predictive Aggregate Transport Model for Microfiltration of Combined Macromolecular Solutions and Poly-Disperse Suspensions: Testing Model with Transgenic Goat MilkBiotechnology Progress, 2003
- A Predictive Aggregate Transport Model for Microfiltration of Combined Macromolecular Solutions and Poly-Disperse Suspensions: Model DevelopmentBiotechnology Progress, 2003
- Contributions to electrostatic interactions on protein transport in membrane systemsAIChE Journal, 2001
- Model Process for Removal of Caseins from Milk of Transgenic AnimalsBiotechnology Progress, 2001
- Electrophoretic analisys to detect and quantify additional whey in milk and dairy beveragesFood Science and Technology, 2000
- Casein sub-micelles: do they exist?International Dairy Journal, 1999
- Production of biologically active salmon calcitonin in the milk of transgenic rabbitsNature Biotechnology, 1998
- High-level expression of recombinant human fibrinogen in the milk of transgenic miceNature Biotechnology, 1996
- The behavior of suspensions and macromolecular solutions in crossflow microfiltrationJournal of Membrane Science, 1994
- Optimization of Solute Separation by DiafiltrationSeparation Science, 1976