Breakage of transgenic tobacco roots for monoclonal antibody release in an ultra-scale down shearing device.
Bottom Line: A possible method for extraction of MAbs from roots is by homogenization, breaking the roots into fragments to release the antibody.Size distributions of the remaining intact roots and root fragments were obtained as a function of shearing time.It was postulated that root breakage in such a high shearing device was due to root-impeller collisions and the particle size data suggest that roots colliding with the impeller were completely fragmented into debris particles of the order of 0.1 mm in length.
Affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.Show MeSH
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Mentions: Interestingly, the distributions of lengths of roots and fragments after all shearing times were bimodal and were dominated in number by the fragments. Figure 1a shows a combined frequency distribution of lengths of roots and fragments after 120 s of shearing, as an example. The distributions strongly suggest breakage of roots was by complete fragmentation of previously intact roots, presumably on collision with the impeller. Figure 1b is an expansion of Figure 1a, for fragments less than or equal to 0.2 mm. The mean fragment length (i.e., excluding intact roots only) for the combined data at all shearing times was 0.135 ± 0.003 mm and the mode was 0.064 mm. It can be inferred that each root broke into 74 fragments on average. By microscopic examination of the roots, the size of a typical cell was determined to be about 100 µm × 40 µm. This and the data in Figure 1 suggest fragmentation was into single cells or small cell aggregates, with accompanying debris.
Affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.