|Why do some entries have more than one light and/or heavy chain sequence?
|We are displaying any returned sequences that meet the threshold for comprising at least 30% of all Amplicon Sequence Variants (ASVs) from that sample. It is now established that monoclonal hybridomas can express more than one light and/or heavy chain (e.g., Bradbury et al., MABS 10: 539, 2018). Moreover, a subset of these samples are not monoclonal but are oligoclonal, as indicated in the category title. Note that the aberrant light chain sequence derived from SP2/0 myeloma cell partner is not shown.
|How are R-mAbs and scFvs evaluated?
|For evaluation of R-mAbs and scFvs cloned using NeuroMabSeq sequences, plasmids were first transfected into mammalian cells. Conditioned culture medium (tissue culture supernatant) was collected from the transfected cell cultures and assayed for immunoreactivity in side-by-side comparisons to the progenitor mAb in various applications. For R-mAbs, these included immunoblots against rat brain samples, immunohistochemistry against rat brain sections, and/or immunocytochemistry against cultured hippocampal neurons or heterologous cells transiently transfected to express the target protein. The same set of assays was performed on scFvs except that immunoblots were not employed. Which of the specific applications were used was guided by the previously established performance characteristics of the progenitor mAb as detailed here and on the respective datasheets for the individual mAbs.
|sequences obtained from the specific subcloned hybridoma cell lines producing the mAbs that comprise the NeuroMab collection (http://neuromab.ucdavis.edu/neuromabs.cfm)
|sequences obtained from subcloned hybridoma cell lines producing other mAbs that passed multiple validation criteria but that are not part of the NeuroMab collection
|NeuroMab Alternative Subclones
|sequences obtained from alternate subcloned hybridoma cell lines producing the mAbs that comprise the NeuroMab collection
|Lead oligoclonal Abs
|sequences obtained from non-subcloned/oligoclonal hybridoma cell lines producing one or more mAbs that passed multiple validation criteria
|Other oligoclonal Abs
|sequences obtained from non-subcloned/oligoclonal hybridoma cell lines producing one or more mAbs that passed one or more validation criteria
|sequences obtained from plasmids expressing validated recombinant mAbs (see Andrews et al., 2019: eLife 8:e43322. PMID: 300667360)
|commercially sequenced: sequences independently obtained by outside vendors from the specific subcloned hybridoma cell lines producing the mAbs that comprise the NeuroMab collection