base assigner

class abstar.assigners.assigner.BaseAssigner(db_name)

BaseAssigner provides an abstract base class for custom VDJ Assigners.

Several different tools exist for inferring the germline components of recombined antibody sequences. Due to their differing design goals, these tools often have different priorities – some are optimized for speed with a small (often negligible) accuracy penalty, while others define recombination junctions with high precision at the cost of speed. Because there isn’t a single germline assignment tool that is optimal for every use case, the germline assignment component of AbStar has been designed to be modular and replaceable, allowing the relatively straightforward addition of new germline assignment tools and the selection of the desired Assigner at runtime. AbStar’s default Assigner (blastn) identifies V- and J-genes using BLASTn and identifies D-genes using rapid Smith-Waterman local alignment.

The basic purpose of any custom Assigner class is to accept sequences and produce VDJ objects. VDJ objects package the input sequence together with GermlineSegment objects that contain information about V, D and/or J assignments. Following germline assignment by the Assigner, addtional annotation will be performed by AbStar. The VDJ objects provide a common interface by which various Assigners can communicate with the additional sequence annotation components in AbStar. This additional annotation is consistent regardless of the Assigner (as is the output schema), and provides a unified method by which different germline Assigners can be used across different projects while maintaining schematic compatibility with downstream analysis tools.

BaseAssigner provides the following attributes:

  • assigned: a list designed to contain VDJ objects for which
    successful V(D)J assignment has been completed. These VDJ objects will be additionally annotated by AbStar following assignment. Exception/log information in these VDJ objects will only be written to the log file if AbStar is run in debug mode.
  • unassigned: a list designed to contain VDJ objects for which
    V(D)J assigjment was unsuccessful. These VDJ objects will not be additionally annotated by AbStar and any exception/log information contained in these VDJ objects will be logged (even if AbStar was not run in debug mode).
  • germline_directory: path to the directory containing germline databases.
    Germline DBs are heirarchically grouped into separate folders first by species, then by the Assigner for which they were designed (blastn, for example). Within each folder, germline DB files are named as {segment}.fasta, so the Blast V-gene DB for human would be human/blast/v.fasta. Note that this will change as AbStar is updated to annotate TCRs in addition to antibodies
  • binary_directory: path to the directory containing Assigner binaries. Binaries
    are named as {binary}_{system}, where system is the lowercase output from platform.system(). For example, the Blastn binary for OSX would be at binary_directory/blastn_darwin. Some assigners require more than a simple binary for execution (Partis, for example). In this case, all required elements will be placed into a folder within the binary directory, named using the same convention as binaries. So the folder containing Linux-compiled Partis components will be binary_directory/partis_linux. The main Partis executable would then be located at binary_directory/partis_linux/bin/partis.

In order to build a custom assignment class, you simply need to subclass BaseAssigner, implement your assigner, and register your assigner in abstar.assigners.registry.ASSIGNERS. Obviously, this is in addition to adding any required binaries and specialized germline database formats. AbStar prefers to incorporate assigner binaries if at all possible (as opposed to relying on system installations) to smooth the process of installing AbStar and eliminate the need for users to install several additional programs before having a fully operational AbStar installation.

There are two critical elements to the design of your assigner class. First, ensure that you call the BaseAssigner’s __init__() at the start of your class’s __init__(). Second, you must implement the __call__() method.

Your custom assigner class’s __init__() must accept a single argument (species, in addition to the required self). You must then call the BaseAssigner’s __init__() and pass the species argument. The simplest was to do this is by using super(), like this: super(MyAssigner, self).__init__(species), where MyAssigner is your assigner class.

  • __call__() must accept two arguments. The first (sequence_file) is a FASTA- or FASTQ-formatted file of input sequences, depending on the user-provided input. The second argument (file_format) defines the format of sequence_file, either 'fasta' or 'fastq'. If a FASTQ file is required by your assigner, ensure that __call__() raises the appropriate exception (and provides a sufficiently clear description of the problem) if a FASTA-formatted file is provided.

  • The result of __call__() should be the generation of a VDJ object for each input sequence (more on this below). The VDJ objects should be appended to either the assigned or unassigned attribute, depending on the outcome of the V(D)J assignment. A high-level overview of how __call__() should work is as follows:

    def __call__(self, seqs, species):
        for seq in seqs:
            vdj = self.assign_vdj(seq, species)
            if self.is_properly_assigned(vdj):

    In this case, the output of assign_vdj() should be a VDJ object.

AbStar defines VDJ objects, which are designed to package the input sequence and GermlineSegment objects that provide information about germline assignment for V, D and/or J genes. VDJ objects must contain the input sequence and may also contain information about V, D and J assignments. While the sequence is required, the germline assignments are optional, to allow creation of VDJ objects for light chains (which lack a D-gene) and/or for unsuccessful assignments. Additionally, this allows flexibility in designing the __call__() method. Assigners can either instantiate a VDJ object at the start of __call__() and attach GermlineSegment``s as they are computed or they can instantiate ``VDJ objects upon completion of V(D)J assignment using the input sequences and computed GermlineSegment``s as instantiation arguments. Because ``VDJ objects contain built-in logging capability, such objects should be created for each input sequence even in the case of unsuccessful assignments so that assignment issues can be logged and reported.

VDJ objects include a logging method: log(). Using log() is similar to using Python’s builtin print() function. Multiple arguments to log() will be space-delimited, and a sep keyword allows you to provide an alternate delimiter. Log messages are typically used to record useful information or to note the occurance of errors/exceptions that are caught and handled by the Assigner. It is also a useful means by which to note the reason a specific input sequence is unable to be fully processed (because it isn’t a valid antibody sequence, etc). By convention, AbStar log entries should contain the name of the log event (in all caps) followed by a colon and additional log information. An example log() call in the case of a V(D)J assignment that was aborted because the V-gene alignment score didn’t meet the alignment quality threshold:

for seq in seqs:
    vdj = VDJ(seq)
    v = assign_v_gene(seq)
    if v.score < threshold:
        vdj.log('V-GENE ASSIGNMENT ERROR:', 'Alignment score ({}) was too low'.format(v.score))

VDJ objects also contain a mechanism for logging exceptions: exception(). This is typically used to log unexpected exceptions (ie, those that are not caught and handled by the Assigner). It is useful to provide some information about when/where in the Assigner the exception occured, as well as a formatted version of the exception traceback. An example of exceptions() being used to log an exception that occurs during V-gene assignment:

vdj = VDJ(seq)
    v = assign_v_gene(seq)
    vdj.exception('V-GENE ASSIGNMENT EXCEPTION:', traceback.format_exc())

One of the primary benefits of recording log and exception information through the VDJ objects is that it allows for much easier formatting of such events when multiple sequences are being processed in parallel. Using Python’s logging utility would result in log events being logged as they occur, which means that multiple log entries for the same input sequence may be far apart in the resulting log file. Packaging log and exception information with each VDJ object alows AbStar to create a much more useful log file in which all information related to a single input sequence is found in a single log location.

GermlineSegment objects contain information about the assigned germline gene segment (V, D or J). Instantiation of a GermlineSegment object requires only the name of the germline gene segment (in IMGT format, like ‘IGHV3-23*01’) and the database name. Several other optional arguments can be included at instantiation. score is the assignment score, typically an int or float. strand indicates the strand orientation of the input sequence ('+' or '-'). others is a list of additional high scoring GermlineSegment objects. assigner_name is the name of the custom Assigner (as a str), and will be converted to lowercase before recording in AbStar output.

GermlineSegment objects also provide a few additional public attributes that may be desirable for certain custom Assigners, especially those that have been specifically designed to accurately define recombination regions. query_start, query_end, germline_start and germline_end allow the Assigner to define the start and end points of the germline alignment for both the query and germline sequences. Note that both query_start and query_start and query_end MUST be relative to the complete query sequence in the '+' orientation (in other words, V-gene at the 5’ end and J-gene at the 3’ end). AbStar realigns all assigned germline genes using alignment parameters designed to accurately identify somatic mutations as well as somatic mutation-induced indels. If the additional assignment position attributes are provided by the Assigner, AbStar will force their use during realignment. If not, AbStar will use the assigned germline genes for realignment and will define the start and end points of the germline alignment during realignment.


The following is an example of a custom Assigner class, named MyAssigner. It uses BioPython’s SeqIO to parse the input file, and because BioPython is a dependency of AbStar, you can assume that BioPython will be present on any system running AbStar. All custom Assigners should be located in the assigners directory:

import os
import platform
import traceback

from abstar.assigners.assigner import BaseAssigner
from abstar.core.vdj import VDJ
from abstar.core.germline import GermlineSegment

from Bio import SeqIO

class MyAssigner(BaseAssigner):

    def __init__(self, db_name):
        super(MyAssigner, self).__init__(db_name)
        self.binary = os.path.join(self.binary_directory, 'mybinary_{}'.format(platform.system()))

    def __call__(self, sequence_file, file_format):
        for sequence in SeqIO.parse(open(sequence_file, 'r'), file_format):
            seq = Sequence(sequence)
            vdj = VDJ(seq)

            # V-gene assignment
            # The VDJ object (rather than just the sequence) is passed to the assignment
            # function for three reasons:
            #    1) it contains the sequence, so we don't need to pass it separately
            #    2) it allows logging during germline assignment, via vdj.log()
            #    3) subsequent assignments will have access to information about
            #       previous assignments (J-gene assignment operations have access
            #       to information about the V-gene assignment, etc.)
                vdj.v = self.assign_germline(vdj, 'V')
                vdj.exception('V-ASSIGNMENT:', traceback.format_exc())

            # J-gene assignment
                vdj.j = self.assign_germline(vdj, 'J')
                vdj.exception('J-ASSIGNMENT:', traceback.format_exc())

            # D-gene assignment
                vdj.d = self.assign_germline(vdj, 'D')
                vdj.exception('D-ASSIGNMENT:', traceback.format_exc())

            return vdj

    def assign_germline(self, vdj, segment):
        germ_db = os.path.join(self.germline_directory, '{}/ungapped/{}.fasta'.format(self.db_name,

        # do stuff to assign the germline gene, using the species-appropriate germline DB
        # ...
        # ...
        # ...
        # germs is a list of germline genes, ordered by score (highest first)

        if germs[0].score < threshold:
            vdj.log('{}-ASSIGNMENT ERROR:'.format(segment),
                    'Score ({}) is too low'.format(germs[0].score))
            return None
        others = [GermlineSegment(, self.db_name, score=germ.score) for germ in germs[1:6]]
        return GermlineSegment(germs[0].name, self.db_name, score=germs[0].score, others=others)