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Monoclonal antibodies are commonly assumed to be monospecific, but anecdotal studies have reported genetic diversity in antibody heavy chain and light chain genes found within individual hybridomas. As the prevalence of such diversity has never been explored, we analyzed 185 random hybridomas, in a large multicenter dataset. The hybridomas analyzed were not biased towards those with cloning difficulties or known to have additional chains. Of the hybridomas we evaluated, 126 (68.1%) contained no additional productive chains, while the remaining 59 (31.9%) contained one or more additional productive heavy or light chains. The expression of additional chains degraded properties of the antibodies, including specificity, binding signal and/or signal-to-noise ratio, as determined by enzyme-linked immunosorbent assay and immunohistochemistry. The most abundant mRNA transcripts found in a hybridoma cell line did not necessarily encode the antibody chains providing the correct specificity. Consequently, when cloning antibody genes, functional validation of all possible VH and VL combinations is required to identify those with the highest affinity and lowest cross-reactivity. These findings, reflecting the current state of hybridomas used in research, reiterate the importance of using sequence-defined recombinant antibodies for research or diagnostic use.

Purpose: The purpose of this article is to describe an automated technique that creates a mirror-image wax model of the contralateral ear using a rapid prototyping technique, which allows precise positioning and adaptation to the deficient side of the face. Materials and Methods: Magnetic resonance images (MRI) are taken of the face and then digitized and reformatted into a 3-dimensional image. The image of the normal ear is extracted, mirrored, and positioned onto the image of the deformed side of the face. The digitized data, when used in conjunction with stereolithography, produces a photopolymerized resin model ear. A model wax ear is created by pouring molten wax into a silicone mold of the resin model ear Results: A model wax ear of identical dimensions, shape, and internal contouring as the contralateral normal ear is created. The technique allows the undersurface of the model wax ear to be precisely positioned against the surface contours of the deformed side of the face in 1 patient- Conclusion: The use of stereolithography from MRI scan data is a suitable technique for producing a model wax ear that fits the deformed side of the face. The 3-dimensional computer image of the face may also be used to plan the position (level and protrusion) of the ear. [ABSTRACT FROM AUTHOR]