Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation

  1. Tino Pleiner
  2. Mark Bates
  3. Sergei Trakhanov
  4. Chung-Tien Lee
  5. Jan Erik Schliep
  6. Hema Chug
  7. Marc Böhning
  8. Holger Stark
  9. Henning Urlaub
  10. Dirk Görlich  Is a corresponding author
  1. Max Planck Institute for Biophysical Chemistry, Germany
  2. University Medical Center Göttingen, Germany

Decision letter

  1. Karsten Weis
    Reviewing Editor; ETH Zürich, Switzerland

In the interests of transparency, eLife includes the editorial decision letter and accompanying author responses. A lightly edited version of the letter sent to the authors after peer review is shown, indicating the most substantive concerns; minor comments are not usually included.

Thank you for submitting your work entitled "Nanobodies: site-specific labeling for superresolution imaging, rapid epitope-mapping & native protein complex isolation" for consideration by eLife. Your article has been reviewed by two peer reviewers, and the evaluation has been overseen by Karsten Weis as Reviewing Editor and Randy Schekman as the Senior Editor.

The reviewers have discussed the reviews with one another and the Reviewing Editor has drafted this decision to help you prepare a revised submission.

Summary:

There was consensus amongst the reviewers that the manuscript by Pleiner et al. is an impressive study that will be of great use to many labs in cell biology and biochemistry describing valuable tools and approaches for the specific study of the nuclear pore complex. The reviewers had no major scientific issues with the study but made multiple suggestions to edit the text. For example, the Abstract was considered to be not as strong or clear as the research deserves. Clarity could be achieved with the rewording of the Abstract to stronger emphasize the scientific achievements. Similar points were made for the main the text, too. See, for example, the fourth paragraph of the subsection “Site-specific fluorescent labeling of nanobodies”.

https://doi.org/10.7554/eLife.11349.015

Author response

There was consensus amongst the reviewers that the manuscript by Pleiner et al. is an impressive study that will be of great use to many labs in cell biology and biochemistry describing valuable tools and approaches for the specific study of the nuclear pore complex. The reviewers had no major scientific issues with the study but made multiple suggestions to edit the text. For example, the Abstract was considered to be not as strong or clear as the research deserves. Clarity could be achieved with the rewording of the Abstract to stronger emphasize the scientific achievements. Similar points were made for the main the text, too. See, for example, the fourth paragraph of the subsection “Site-specific fluorescent labeling of nanobodies”.

We accommodated all suggested improvements to the Abstract and main text, added the requested experimental details, as well as the PDB accession code (5E0Q) for the anti-Nup98 nanobody TP377 - Nup98 crystal structure. The only suggestion we did not implement was to move Figure 5D to the supplements, because we still believe that this is a very central piece of data (showing that maleimide labeling of nanobodies works through at least 6 different ectopic cysteine positions).

https://doi.org/10.7554/eLife.11349.016

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  1. Tino Pleiner
  2. Mark Bates
  3. Sergei Trakhanov
  4. Chung-Tien Lee
  5. Jan Erik Schliep
  6. Hema Chug
  7. Marc Böhning
  8. Holger Stark
  9. Henning Urlaub
  10. Dirk Görlich
(2015)
Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation
eLife 4:e11349.
https://doi.org/10.7554/eLife.11349

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https://doi.org/10.7554/eLife.11349