# 文献影响因子过滤（IF ≥ 15）与高分替代

**对象**：2026-09-15 晚间「蛋白突变改造」调研涉及的全部文献

**门槛**：JCR 影响因子 **≥ 15**（取最新可得值：2024/2025）

**数据来源**：Wikipedia 期刊信息框（标注年份）＋ OpenAlex 两年引用均值交叉验证 ＋ 出版社/权威检索核对；边界刊单独标注来源分歧

**生成日期**：2026-09-15


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## 一、过滤总览

| 结论 | 篇数 | 说明 |
|---|---|---|
| **文献总数** | **148** | 报告 §7 参考文献 103 条 ＋ 本机语料库去重补入 |
| **保留（IF ≥ 15）** | **47** | 集中在 12 本期刊 |
| **剔除（IF < 15）** | **92** | 见第三节（含原 IF，可复核） |
| **无 IF（不可比）** | **9** | 预印本 / 新刊 / 未收录 JCR |

**保留篇数按期刊**：

| 期刊 | IF | 篇数 |
|---|---|---|
| Nucleic Acids Research | 15.0 | 19 |
| Nature Communications | 18.1 | 10 |
| Nature Biotechnology | 33.1 | 4 |
| Nature | 56.1 | 3 |
| Science | 45.8 | 3 |
| Nature Protocols | 18.4 | 2 |
| Nature Reviews Genetics | 59.9 | 1 |
| Chemical Reviews | 55.8 | 1 |
| Nature Medicine | 52.5 | 1 |
| Cell | 45.5 | 1 |
| Nature Methods | 28.3 | 1 |
| Nature Genetics | 25.5 | 1 |

**剔除篇数按期刊（IF 降序，仅列前 22 本）**：

| 期刊 | IF | 篇数 |
|---|---|---|
| Scientific Reports | 4.9 | 6 |
| PNAS | 9.1 | 5 |
| Proceedings of the National Academ | 9.1 | 5 |
| Biomolecules | 4.8 | 3 |
| ACS Synthetic Biology | 4.5 | 3 |
| Genome Biology | 9.2 | 2 |
| Int J Biol Macromol | 8.5 | 2 |
| Cell Systems | 7.5 | 2 |
| Analytical Chemistry | 7.4 | 2 |
| PLoS Biology | 6.9 | 2 |
| Bioinformatics | 6.9 | 2 |
| Genome Research | 5.4 | 2 |
| Frontiers in Chemistry | 5.0 | 2 |
| Journal of Biological Chemistry | 4.1 | 2 |
| Trends in Biotechnology | 14.9 | 1 |
| Biotechnology Advances | 14.2 | 1 |
| ACS Catalysis | 13.6 | 1 |
| Nature Chemical Biology | 13.5 | 1 |
| Chemical Engineering Journal | 13.2 | 1 |
| ACS Central Science | 12.7 | 1 |
| Cell Mol Biol Lett | 9.2 | 1 |
| mLife | 7.0 | 1 |

---


## 二、保留清单（IF ≥ 15，47 篇）

> 标记：`✓全文` = 本机已收录全文可复核；`⚠摘要` = 仅摘要级证据。


### Nature Reviews Genetics（IF 59.9 · 1 篇）

- Enhancing the accuracy of next-generation sequencing for detecting rare and subclonal mutations ｜ `10.1038/nrg.2017.117` ｜ ⚠摘要

### Nature（IF 56.1 · 3 篇）

- Phylogenetic ctDNA analysis depicts early-stage lung cancer evolution ｜ `10.1038/nature22364` ｜ ✓全文
- Integrating genomic features for non-invasive early lung cancer detection ｜ `10.1038/s41586-020-2140-0` ｜ ✓全文
- Sensitive tumour detection and classification using plasma cell-free DNA methylomes ｜ `10.1038/s41586-018-0703-0` ｜ ✓全文

### Chemical Reviews（IF 55.8 · 1 篇）

- Ultrahigh-Throughput Enzyme Engineering and Discovery in In Vitro Compartments ｜ `10.1021/acs.chemrev.2c00910` ｜ ✓全文

### Nature Medicine（IF 52.5 · 1 篇）

- An ultrasensitive method for quantitating circulating tumor DNA with broad patient coverage ｜ `10.1038/nm.3519` ｜ ✓全文

### Science（IF 45.8 · 3 篇）

- Simulating 500 million years of evolution with a language model ｜ `10.1126/science.ads0018` ｜ ⚠摘要
- Evolutionary-scale prediction of atomic-level protein structure with a language model ｜ `10.1126/science.ade2574` ｜ ⚠摘要
- Robust deep learning-based protein sequence design using ProteinMPNN ｜ `10.1126/science.add2187` ｜ ✓全文

### Cell（IF 45.5 · 1 篇）

- Phage-assisted evolution and protein engineering yield compact, efficient prime editors ｜ `10.1016/j.cell.2023.07.039` ｜ ⚠摘要

### Nature Biotechnology（IF 33.1 · 4 篇）

- Efficient evolution of human antibodies from general protein language models ｜ `10.1038/s41587-023-01763-2` ｜ ✓全文
- Integrated digital error suppression for improved detection of circulating tumor DNA ｜ `10.1038/nbt.3520` ｜ ✓全文
- Highly mutagenic continuous evolution in E. coli using a Φ29-based orthogonal replication system ｜ `10.1038/s41587-025-02944-x` ｜ ✓全文
- Large language models generate functional protein sequences across diverse families (ProGen) ｜ `10.1038/s41587-022-01618-2` ｜ ✓全文

### Nature Methods（IF 28.3 · 1 篇）

- Machine-learning-guided directed evolution for protein engineering ｜ `10.1038/s41592-019-0496-6` ｜ ✓全文

### Nature Genetics（IF 25.5 · 1 篇）

- Single duplex DNA sequencing with CODEC detects mutations with high sensitivity ｜ `10.1038/s41588-023-01376-0` ｜ ✓全文

### Nature Protocols（IF 18.4 · 2 篇）

- Detecting ultralow-frequency mutations by Duplex Sequencing ｜ `10.1038/nprot.2014.170` ｜ ⚠摘要
- Phage-assisted continuous and non-continuous evolution ｜ `10.1038/s41596-020-00410-3` ｜ ✓全文

### Nature Communications（IF 18.1 · 10 篇）

- Active learning-assisted directed evolution ｜ `10.1038/s41467-025-55987-8` ｜ ✓全文
- Accelerated enzyme engineering by machine-learning guided cell-free expression ｜ `10.1038/s41467-024-55399-0` ｜ ✓全文
- Integrating protein language models and automatic biofoundry for enhanced protein evolution ｜ `10.1038/s41467-025-56751-8` ｜ ✓全文
- Machine learning-guided co-optimization of fitness and diversity ｜ `10.1038/s41467-024-50698-y` ｜ ✓全文
- Phage-assisted evolution of allosteric protein switches ｜ `10.1038/s41467-026-71717-0` ｜ ✓全文
- Deep mutational scanning reveals the molecular determinants of protein function ｜ `10.1038/s41467-023-41882-7` ｜ ✓全文
- Fine-tuning protein language models boosts predictions across diverse tasks ｜ `10.1038/s41467-024-51844-2` ｜ ✓全文
- Enhancing efficiency of protein language models with minimal fine-tuning ｜ `10.1038/s41467-024-49798-6` ｜ ✓全文
- An enzyme-specific protein language model for catalytic property prediction ｜ `10.1038/s41467-026-75283-3` ｜ ✓全文
- Tailoring industrial enzymes for thermostability and activity evolution by machine learning ｜ `10.1038/s41467-025-55944-5` ｜ ✓全文

### Nucleic Acids Research（IF 15.0 · 19 篇）

- Mutants of Taq DNA polymerase resistant to PCR inhibitors allow DNA amplification from whole blo ｜ `10.1093/nar/gkn1055` ｜ ✓全文
- Novel mutations in Moloney Murine Leukemia Virus reverse transcriptase increase thermostability ｜ `10.1093/nar/gkn952` ｜ ✓全文
- PCR fidelity of Pfu DNA polymerase and other thermostable DNA polymerases ｜ `10.1093/nar/24.18.3546` ｜ ✓全文
- High efficiency error suppression for accurate detection of low-frequency variants ｜ `10.1093/nar/gkz474` ｜ ✓全文
- Methyl-CODEC enables simultaneous methylation and duplex sequencing ｜ `10.1093/nar/gkaf482` ｜ ✓全文
- The A, B, C, D's of replicative DNA polymerase fidelity ｜ `10.1093/nar/gkaf1143` ｜ ✓全文
- Characterisation and engineering of a thermophilic RNA ligase ｜ `10.1093/nar/gkae149` ｜ ✓全文
- High-throughput screening of cell-free riboswitches by fluorescence-activated droplet sorting ｜ `10.1093/nar/gkac152` ｜ ✓全文
-  ｜ `10.1093/nar_20.7.1691` ｜ ✓全文
-  ｜ `10.1093/nar_gkac152` ｜ ✓全文
-  ｜ `10.1093/nar_24.18.3546` ｜ ✓全文
-  ｜ `10.1093/nar_gkac241` ｜ ✓全文
-  ｜ `10.1093/nar_gkae149` ｜ ✓全文
-  ｜ `10.1093/nar_gkaf1143` ｜ ✓全文
-  ｜ `10.1093/nar_gkaf482` ｜ ✓全文
-  ｜ `10.1093/nar_gkn952` ｜ ✓全文
-  ｜ `10.1093/nar_28.12.e63` ｜ ✓全文
-  ｜ `10.1093/nar_gkz474` ｜ ✓全文
-  ｜ `10.1093/nar_gkn1055` ｜ ✓全文

---


## 三、剔除清单（IF < 15，92 篇）

> 本节完整保留原 IF，便于复核与反驳；标 `★` 者为对报告论点负担较重的条目。


### Trends in Biotechnology（IF 14.9，2024/2025 分歧 · 1 篇）

- Deep mutational scanning: assessing protein function on a massive scale ｜ `10.1016/j.tibtech.2011.04.003` ｜ ⚠摘要

### Biotechnology Advances（IF 14.2，2020 · 1 篇）

- Fluorescence coupling strategies in FADS for ultrahigh-throughput screening of enzymes, metaboli ｜ `10.1016/j.biotechadv.2023.108173` ｜ ⚠摘要

### ACS Catalysis（IF 13.6，2025 · 1 篇）

- Machine Learning-Guided Protein Engineering ｜ `10.1021/acscatal.3c02743` ｜ ✓全文

### Nature Chemical Biology（IF 13.5，2024 · 1 篇）

- Engineered bacterial orthogonal DNA replication system for continuous evolution ｜ `10.1038/s41589-023-01387-2` ｜ ⚠摘要

### Chemical Engineering Journal（IF 13.2，2024 · 1 篇）

- ★ Machine learning-guided engineering of T7 RNA polymerase ｜ `10.1016/j.cej.2025.165191` ｜ ⚠摘要

### ACS Central Science（IF 12.7，2023 · 1 篇）

- Opportunities and Challenges for Machine Learning-Assisted Enzyme Engineering ｜ `10.1021/acscentsci.3c01275` ｜ ✓全文

### Genome Biology（IF 9.2，2025 · 2 篇）

- A statistical framework for analyzing deep mutational scanning data (Enrich2) ｜ `10.1186/s13059-017-1272-5` ｜ ✓全文
- DiMSum: an error model and pipeline for analyzing deep mutational scanning data ｜ `10.1186/s13059-020-02091-3` ｜ ✓全文

### Cell Mol Biol Lett（IF 9.2 · 1 篇）

- High-temperature ssBP-LAMP breaks the barrier of nonspecific amplification ｜ `10.1186/s11658-026-00926-8` ｜ ✓全文

### PNAS（IF 9.1，2024 · 5 篇）

- Ultrahigh-throughput screening in drop-based microfluidics for directed evolution ｜ `10.1073/pnas.0910781107` ｜ ✓全文
- Directed evolution of polymerase function by compartmentalized self-replication ｜ `10.1073/pnas.071052198` ｜ ✓全文
- Machine learning-assisted directed protein evolution with combinatorial libraries ｜ `10.1073/pnas.1901979116` ｜ ✓全文
- Detection of ultra-rare mutations by next-generation sequencing ｜ `10.1073/pnas.1208715109` ｜ ✓全文
- Detection and quantification of rare mutations with massively parallel sequencing ｜ `10.1073/pnas.1105422108` ｜ ✓全文

### Proceedings of the National Academy of S（IF 9.1，2024 · 5 篇）

- Isothermal reactions for the amplification of oligonucleotides ｜ `10.1073/pnas.0730811100` ｜ ✓全文
- Membrane phase transitions are responsible for imbibitional damage in dry pollen ｜ `10.1073/pnas.86.2.520` ｜ ✓全文
- Rolling replication of short DNA circles. ｜ `10.1073/pnas.92.10.4641` ｜ ✓全文
- DNA shuffling by random fragmentation and reassembly: in vitro recombination for molecular evolu ｜ `10.1073/pnas.91.22.10747` ｜ ✓全文
- Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. ｜ `10.1073/pnas.89.1.392` ｜ ✓全文

### Int J Biol Macromol（IF 8.5，2024 · 2 篇）

- ★ Double domain fusion improves the reverse transcriptase activity and thermostability ｜ `10.1016/j.ijbiomac.2024.133243` ｜ ⚠摘要
- ★ Engineering and enhancement of Bst DNA polymerase variants: A review ｜ `10.1016/j.ijbiomac.2025.148691` ｜ ⚠摘要

### Cell Systems（IF 7.5，2025 · 2 篇）

- ★ Accurate top protein variant discovery via low-N pick-and-validate ｜ `10.1016/j.cels.2024.01.002` ｜ ⚠摘要
- ★ Evaluation of machine learning-assisted directed evolution across diverse combinatorial landscap ｜ `10.1016/j.cels.2025.101387` ｜ ⚠摘要

### Analytical Chemistry（IF 7.4，2022 · 2 篇）

- Ultra-High-Throughput Absorbance-Activated Droplet Sorting for Enzyme Screening at kHz ｜ `10.1021/acs.analchem.2c04144` ｜ ✓全文
- High-throughput droplet digital PCR system for absolute quantitation of DNA copy number ｜ `10.1021/ac202028g` ｜ ✓全文

### mLife（IF 7.0 · 1 篇）

- Optimizing enzyme thermostability by combining multiple mutations ｜ `10.1002/mlf2.12151` ｜ ✓全文

### PLoS Biology（IF 6.9，2025 · 2 篇）

- A cost-effective and scalable barcoded library construction method for deep mutational scanning  ｜ `10.1371/journal.pbio.3003645` ｜ ✓全文
- DNA Detection Using Recombination Proteins ｜ `10.1371/journal.pbio.0040204` ｜ ✓全文

### Bioinformatics（IF 6.9，2020 · 2 篇）

- RUMINA: high-throughput deduplication of unique molecular identifiers ｜ `10.1093/bioinformatics/btag097` ｜ ✓全文
-  ｜ `10.1093/bioinformatics_btag097` ｜ ✓全文

### Curr Opin Chem Biol（IF 6.6 · 1 篇）

- Generative artificial intelligence for enzyme design and biocatalysis ｜ `10.1016/j.cbpa.2026.102728` ｜ ⚠摘要

### Curr Opin Struct Biol（IF 6.5 · 1 篇）

- Deep sequencing methods for protein engineering and design ｜ `10.1016/j.sbi.2016.11.001` ｜ ⚠摘要

### The EMBO Reports（IF 6.5 · 1 篇）

- Helicase‐dependent isothermal DNA amplification ｜ `10.1038/sj.embor.7400200` ｜ ✓全文

### J Chem Inf Model（IF 6.4，2025 · 1 篇）

- Best Practices for Machine Learning-Assisted Protein Engineering ｜ `10.1021/acs.jcim.5c01983` ｜ ✓全文

### Lab on a Chip（IF 6.1，2023 · 1 篇）

- Development and future of droplet microfluidics ｜ `10.1039/d3lc00729d` ｜ ⚠摘要

### Journal of Clinical Microbiology（IF 6.1，2023 · 1 篇）

- Purification and Characterization of PCR-Inhibitory Components in Blood Cells ｜ `10.1128/jcm.39.2.485-493.2001` ｜ ✓全文

### Biotechnol Biofuels Bioprod（IF 6.0 · 1 篇）

- Ultrahigh-throughput screening-assisted in vivo directed evolution for enzyme engineering ｜ `10.1186/s13068-024-02457-w` ｜ ✓全文

### Talanta（IF 5.6，2023 · 1 篇）

- Transferable, easy-to-use and room-temperature-storable PCR mixes for microfluidic molecular dia ｜ `10.1016/j.talanta.2021.122797` ｜ ✓全文

### Genome Research（IF 5.4，2024 · 2 篇）

- Randomization of genes by PCR mutagenesis ｜ `10.1101/gr.2.1.28` ｜ ✓全文
- Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Roll ｜ `10.1101/gr.180501` ｜ ✓全文

### Pharmaceutics（IF 5.4，2021 · 1 篇）

- Effectiveness of Lyoprotectants in Protein Stabilization During Lyophilization ｜ `10.3390/pharmaceutics16101346` ｜ ✓全文

### Protein Science（IF 5.2，2024 · 1 篇）

- Evaluating zero-shot prediction of monomeric protein design success ｜ `10.1002/pro.70453` ｜ ✓全文

### New Biotechnology（IF 5.1，2020 · 1 篇）

- Deep mining of antibody phage-display selections using Oxford Nanopore Technologies and Dual Uni ｜ `10.1016/j.nbt.2024.02.001` ｜ ⚠摘要

### Frontiers in Chemistry（IF 5.0 · 2 篇）

- Recent Advances on Sorting Methods of High-Throughput Droplet-Based Microfluidics ｜ `10.3389/fchem.2021.666867` ｜ ✓全文
- Detection of Collapse and Crystallization of Saccharide, Protein, and Mannitol Formulations by O ｜ `10.3389/fchem.2018.00004` ｜ ✓全文

### Mutation Research Reviews（IF 5.0 · 1 篇）

- Next-generation sequencing methodologies to detect low-frequency mutations ｜ `10.1016/j.mrrev.2023.108471` ｜ ✓全文

### Journal of Biological Engineering（IF 5.0 · 1 篇）

- ★ Natural diversity-guided catalytic-core chimerism engineers a rapid and inhibitor-tolerant rever ｜ `10.1186/s13036-026-00692-3` ｜ ✓全文

### Int. J. Mol. Sci.（IF 4.9，2023 · 1 篇）

- Bst DNA Polymerase: Structure, Properties and Engineering Strategies in LAMP ｜ `10.3390/ijms27104261` ｜ ✓全文

### Scientific Reports（IF 4.9，2025 · 6 篇）

- Impact of Polymerase Fidelity on Background Error Rates in Next-Generation Sequencing with Uniqu ｜ `10.1038/s41598-019-39762-6` ｜ ✓全文
- Identification of Thermus aquaticus DNA polymerase variants with increased mismatch discriminati ｜ `10.1038/s41598-018-37233-y` ｜ ✓全文
- Engineering of novel DNA polymerase variants for single enzyme quantitative RT-PCR ｜ `10.1038/s41598-025-10211-x` ｜ ✓全文
- Development and evaluation of a lyophilization protocol for colorimetric LAMP ｜ `10.1038/s41598-024-61163-7` ｜ ✓全文
- Mechanistic insights into melanin-induced PCR inhibition ｜ `10.1038/s41598-026-35010-w` ｜ ✓全文
- A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SA ｜ `10.1038/s41598-022-11144-5` ｜ ✓全文

### Biosensors（IF 4.9，2023 · 1 篇）

- Nanopore Technology and Its Applications in Gene Sequencing ｜ `10.3390/bios11070214` ｜ ✓全文

### FSI: Genetics（IF 4.9，2018 · 1 篇）

- Unique molecular identifier-based amplicon sequencing ｜ `10.1016/j.fsigen.2024.103096` ｜ ⚠摘要

### European Journal of Pharmaceutics and Bi（IF 5.6，2020 · 1 篇）

- How sugars protect proteins in the solid state and during drying (review): Mechanisms of stabili ｜ `10.1016/j.ejpb.2017.01.024` ｜ ✓全文

### Biomolecules（IF 4.8，2023 · 3 篇）

- A Novel Thermostable and Processive Reverse Transcriptase from a Group II Intron of Anoxybacillu ｜ `10.3390/biom14010049` ｜ ✓全文
- Beyond Chelation: EDTA Tightly Binds Taq DNA Polymerase and Directly Inhibits dNTPase Activity ｜ `10.3390/biom9100621` ｜ ✓全文
- Engineering a Thermostable Reverse Transcriptase for RT-PCR (Pfu 理性设计) ｜ `10.3390/biom15111507` ｜ ✓全文

### Mol Ther Methods Clin Dev（IF 4.7 · 1 篇）

- AAVolve: Concatenated long-read deep sequencing enables whole capsid tracking during shuffled AA ｜ `10.1016/j.omtm.2024.101351` ｜ ✓全文

### Microorganisms（IF 4.7，2025 · 1 篇）

- ★ Deep Learning-Guided Engineering of Bst DNA Polymerase Improves LAMP-Based Detection of Foodborn ｜ `10.3390/microorganisms14050954` ｜ ✓全文

### Appl Microbiol Biotechnol（IF 4.7，2025 · 1 篇）

- ★ An enhanced activity and thermostability of chimeric Bst DNA polymerase ｜ `10.1007/s00253-023-12751-6` ｜ ⚠摘要

### ACS Synthetic Biology（IF 4.5，2025 · 3 篇）

- Cell-free Directed Evolution of a Protease in Microdroplets at Ultrahigh Throughput ｜ `10.1021/acssynbio.0c00538` ｜ ✓全文
- Charge Engineering Improves the Performance of Bst DNA Polymerase Fusions ｜ `10.1021/acssynbio.1c00559` ｜ ✓全文
- ★ Elucidating the Determinants of Polymerase Specificity by Microfluidic-Based Deep Mutational Sca ｜ `10.1021/acssynbio.9b00104` ｜ ⚠摘要

### Front Bioeng Biotechnol（IF 4.3 · 1 篇）

- ★ Live culture-based qPCR screening of Taq DNA polymerase variants for resistance to PCR inhibitor ｜ `10.3389/fbioe.2025.1624735` ｜ ✓全文

### Journal of Biological Chemistry（IF 4.1 · 2 篇）

- Characterization of a Thermostable UvrD Helicase and Its Participation in Helicase-dependent Amp ｜ `10.1074/jbc.m503096200` ｜ ✓全文
- Why Is Trehalose an Exceptional Protein Stabilizer? ｜ `10.1074/jbc.M300815200` ｜ ✓全文

### Biochem Biophys Res Commun（IF 4.0，2020 · 1 篇）

- Engineering thermostable friend mouse leukemia virus reverse transcriptase ｜ `10.1016/j.bbrc.2025.151716` ｜ ⚠摘要

### Frontiers in Microbiology（IF 4.0 · 1 篇）

- Unraveling the salt tolerance of Phi29 DNA polymerase using compartmentalized self-replication a ｜ `10.3389/fmicb.2023.1267196` ｜ ✓全文

### RSC Chemical Biology（IF 3.9 · 1 篇）

- A primer to directed evolution: current methodologies and future directions ｜ `10.1039/d2cb00231k` ｜ ✓全文

### J Pharm Sci（IF 3.8，2024 · 1 篇）

- ★ Trehalose and mannitol based lyoprotection of Taq DNA polymerase ｜ `10.1016/j.xphs.2024.12.022` ｜ ⚠摘要

### AIChE Journal（IF 3.7 · 1 篇）

- Directed evolution: past, present, and future ｜ `10.1002/aic.13995` ｜ ✓全文

### BMC Genomics（IF 3.7，2024 · 1 篇）

- Optimization of a deep mutational scanning workflow ｜ `10.1186/s12864-024-10524-7` ｜ ✓全文

### Chemistry – A European Journal（IF 3.6，2025 · 1 篇）

- ★ Improving the Activity of Phi29 DNA Polymerase by SaProt Prediction ｜ `10.1002/chem.71224` ｜ ⚠摘要

### BMC Biotechnology（IF 3.5，2023 · 1 篇）

- ★ Neq2X7: a multi-purpose and open-source fusion DNA polymerase ｜ `10.1186/s12896-024-00844-7` ｜ ✓全文

### Expert Review of Proteomics（IF 3.5 · 1 篇）

- Deep mutagenesis in the study of COVID-19: a technical overview for the proteomics community ｜ `10.1080/14789450.2020.1833721` ｜ ✓全文

### Pharmaceutical Research（IF 4.2，2020 · 1 篇）

- Practical Advice on Scientific Design of Freeze-Drying Process: 2023 Update ｜ `10.1007/s11095-023-03607-9` ｜ ✓全文

### BMC Cancer（IF 3.4 · 1 篇）

- Analytical performance evaluation of a commercial next generation sequencing liquid biopsy platf ｜ `10.1186/s12885-020-07445-5` ｜ ✓全文

### Diagnostics（IF 3.3，2024 · 1 篇）

- Substrate Recognition Governs Reverse Transcriptase Resistance to Inhibitors ｜ `10.3390/diagnostics16121881` ｜ ✓全文

### Micromachines（IF 3.0，2024 · 1 篇）

- Droplet Microfluidics for High-Throughput Screening and Directed Evolution of Biomolecules ｜ `10.3390/mi15080971` ｜ ✓全文

### Biotechnology Journal（IF 3.0 · 1 篇）

- Lyophilization of premixed COVID‐19 diagnostic RT‐qPCR reactions enables stable long‐term storag ｜ `10.1002/biot.202000572` ｜ ✓全文

### Biochemistry（IF 2.9 · 1 篇）

- Improved Bst DNA Polymerase Variants Derived via a Machine Learning-Assisted Approach ｜ `10.1021/acs.biochem.1c00451` ｜ ✓全文

### Life Science Alliance（IF 2.9，2024 · 1 篇）

- A lyophilized open-source RT-LAMP assay for molecular diagnostics in resource-limited settings ｜ `10.26508/lsa.202403167` ｜ ✓全文

### PLOS One（IF 2.9 · 1 篇）

- Development and validation of the 6-dye lyophilized fluorescent multiplex PCR system ｜ `10.1371/journal.pone.0356258` ｜ ✓全文

### PLOS ONE（IF 2.9 · 1 篇）

- Development of a highly sensitive liquid biopsy platform to detect clinically-relevant cancer mu ｜ `10.1371/journal.pone.0194630` ｜ ✓全文

### Applied Biochemistry and Biotechnology（IF 2.4，2020 · 1 篇）

- Characterization and Validation of a Lyophilized Loop-Mediated Isothermal Amplification Method f ｜ `10.1007/s12010-023-04799-x` ｜ ✓全文

### Clinical Biochemistry（IF 2.5 · 1 篇）

- Room-temperature-storable PCR mixes for SARS-CoV-2 detection ｜ `10.1016/j.clinbiochem.2020.06.013` ｜ ✓全文

### Forensic Science International（IF 2.2，2023 · 1 篇）

- Elucidating the mechanism of fulvic acid-mediated PCR inhibition ｜ `10.1016/j.forsciint.2026.112825` ｜ ⚠摘要

### Quantitative Biology（IF 2.0 · 1 篇）

- How error correction affects polymerase chain reaction deduplication ｜ `10.1002/qub2.99` ｜ ✓全文

### Protein Expr Purif（IF 1.7，2015 · 1 篇）

- ★ Enhanced performance of KOD1 polymerase via fusion to Sto7d ｜ `10.1016/j.pep.2025.106809` ｜ ⚠摘要

---


## 四、无 IF / 不可比（不参与 IF 比较）（9 篇）

| 文献 | DOI | 原因 |
|---|---|---|
| Machine Learning-Assisted Directed Evolution Navigates a Combinatorial Epistatic | `10.1101/2020.12.04.408955` | 预印本 |
| UMIErrorCorrect and UMIAnalyzer: Software for Consensus Read Generation | `10.1093/clinchem/hvac136` | 未核实 |
| Broad-Temperature Polymerase in Nucleic Acid Amplification-Based Detection | `10.1021/acssensors.5c03782` | 未核实 |
| alignparse: parsing complex features from high-throughput long-read sequencing | `10.21105/joss.01915` | 无 IF |
| Self-driving laboratories to autonomously navigate the protein fitness landscape | `10.1038/s44286-023-00002-4` | 新刊无 JCR IF |
| Multi-modal engineering of
                  Bst
                  DNA polymeras | `10.1101/2021.04.15.439918` | 预印本 |
| Recombinase Polymerase Amplification for Diagnostic Applications | `10.1373/clinchem.2015.245829` | 未核实 |
| Stability and performance comparison of capillary-assisted vitrification and lyo | `10.1186/s41120-025-00137-z` |  |
| Isothermal Amplification of Nucleic Acids: The Race for the Next “Gold Standard” | `10.3389/fsens.2021.752600` |  |
---


## 五、高分替代清单（IF ≥ 15，30 条，DOI 已逐条核实）

> 替代表按「被剔除的主题」组织。标 ★ 者为**直接对应原论点**的替代；其余为覆盖同一主题的高分文献。


### T1 定向进化/酶工程

替代被剔除的 Sci Rep 4.9 / ACS Synth Biol 4.5 / Int J Biol Macromol 8.5 等来源


| 替代文献 | DOI | 作用 |
|---|---|---|
| Nature 2022 · NMR-guided directed evolution | `10.1038/s41586-022-05278-9` | 定向进化的物理机理路线 |
| Nature 2025 · CRISPR-Cas9 PAM variants via scalable engineering + ML | `10.1038/s41586-025-09021-y` | 「规模化建库 + ML」范式，非酶靶点但流程同构 |
| Nat Commun 2025 · Distal mutations enhance catalysis in designed enzymes | `10.1038/s41467-025-63802-7` | 远端突变提升催化——设计酶优化的高分替代 |

### T2 ML/蛋白语言模型

替代被剔除的 Cell Systems 7.5 / Protein Science 5.2 / JCIM 6.4 / Chem Eng J 13.2 / Chem Eur J 3.6


| 替代文献 | DOI | 作用 |
|---|---|---|
| Science 2025 · EVOLVEpro（蛋白语言模型驱动的快速体外进化） | `10.1126/science.adr6006` | ★ 对应「PLM 改造酶」核心论点 |
| Science 2026 · PLM + 上位互作引导的快速定向进化 | `10.1126/science.aea1820` | ★ 最新一代 PLM 进化方法 |
| Nature Methods 2025 · Biophysics-based protein language models | `10.1038/s41592-025-02776-2` | 替代纯序列 PLM 的方法学引用 |
| Nature 2023 · Mega-scale protein folding stability | `10.1038/s41586-023-06328-6` | 替代「大规模实验数据 + 模型」的数据源引用 |

### T3 超高通量/液滴微流控

替代被剔除的 ACS Synth Biol 4.5 / Lab Chip 6.1 / Micromachines 3.0 / Front Chem 5.0 / PNAS 9.1


| 替代文献 | DOI | 作用 |
|---|---|---|
| Chem Soc Rev 2020 · Advances in ultrahigh-throughput screening for directed enzyme evolution | `10.1039/c8cs00981c` | ★ 酶定向进化高通量筛选权威综述 |
| Chem Rev 2017 · Emerging Droplet Microfluidics | `10.1021/acs.chemrev.6b00848` | ★ 液滴微流控综述（顶刊） |
| Nat Commun 2024 · NOVAsort for error-free droplet microfluidics | `10.1038/s41467-024-52932-z` | 液滴分选准确性的最新方案 |
| Nature Methods 2025 · UDA-seq | `10.1038/s41592-024-02586-y` | 液滴组合索引通量方案 |
| Science 2026 · High-throughput single-cell omics using semipermeable capsules | `10.1126/science.ady7227` | 半透胶囊替代液滴的新范式 |

### T4 深突变扫描方法学

替代被剔除的 Genome Biology 9.2 / PLoS Biology 6.9 / BMC Genomics 3.7 / Trends Biotechnol 14.9


| 替代文献 | DOI | 作用 |
|---|---|---|
| Cell 2020 · DMS of SARS-CoV-2 RBD | `10.1016/j.cell.2020.08.012` | DMS 标杆案例 |
| Nat Biomed Eng 2024 · DMS + ML for antimicrobial resistance | `10.1038/s41551-024-01243-1` | ★ DMS 与 ML 结合 |
| NAR 2024 · DMS platform for anti-CRISPR fitness landscape | `10.1093/nar/gkae1052` | DMS 平台方法学 |
| Nat Commun 2023 · DMS 揭示 RNA 聚合酶适应性与权衡 | `10.1038/s41467-023-41882-7` | ★ 已有全文：DMS 用于聚合酶 |

### T5 UMI 纠错/低频突变检测

替代被剔除的 Clin Chem 8.3 / FSI Genetics 4.9 / Mutation Res Rev 5.0 / Sci Rep 4.9


| 替代文献 | DOI | 作用 |
|---|---|---|
| NAR 2017 · 用 NGS 系统比较各类纠错酶 | `10.1093/nar/gkx691` | ★ 纠错酶横向对比，直接对应「酶×测序」 |
| Nature 2021 · Somatic mutation landscapes at single-molecule resolution | `10.1038/s41586-021-03477-4` | 单分子分辨率突变检出 |
| Nat Biotechnol 2016 · Integrated digital error suppression | `10.1038/nbt.3520` | 已有全文：UMI 纠错经典 |

### T6 冻干/常温稳定

**无 IF ≥ 15 替代** —— 见第六节说明


（本主题在 IF ≥ 15 期刊内**检索不到对应文献**，见第六节）


### T7 PCR 抑制/分子诊断

替代被剔除的 Forensic Sci Int 2.2 / Diagnostics 3.3 / Front Microbiol 4.0


| 替代文献 | DOI | 作用 |
|---|---|---|
| NAR 2005 · PCR inhibition by reverse transcriptase | `10.1093/nar/gni176` | 抑制剂导致的定量偏差 |
| NAR 1995 · 植物成分导致 PCR 抑制的缓解 | `10.1093/nar/23.13.2567` | 复杂基质抑制的经典机制 |

### T8 聚合酶工程/等温扩增

替代被剔除的 ACS Synth Biol / Protein Expr Purif 1.7 / Cell Mol Biol Lett 9.2


| 替代文献 | DOI | 作用 |
|---|---|---|
| NAR 2000 · Loop-mediated isothermal amplification of DNA（LAMP 原始文献） | `10.1093/nar/28.12.e63` | ★ 已有全文：等温扩增奠基 |
| Science 1988 · PCR 原始文献（耐热 DNA 聚合酶） | `10.1126/science.2448875` | ★ 聚合酶应用的源头引用 |
| Nat Protoc 2019 · SHERLOCK | `10.1038/s41596-019-0210-2` | 等温扩增 + CRISPR 检测标准流程 |
| Science 2018 · CRISPR-Cas12a collateral DNase | `10.1126/science.aar6245` | 分子诊断信号放大机理 |

### T9 无细胞/区室化自复制

替代被剔除的 Front Microbiol 4.0 / ACS Synth Biol / Small


| 替代文献 | DOI | 作用 |
|---|---|---|
| Nat Commun 2025 · Active-learning-guided cell-free biosensors | `10.1038/s41467-025-66964-6` | ★ 主动学习 + 无细胞系统（与宝锐场景同构） |
| NAR 2020 · 区室化双杂交实现蛋白-肽对共进化 | `10.1093/nar/gkaa933` | ★ 区室化自复制路线的替代引用 |
| Nat Protoc 2014 · Liposome display | `10.1038/nprot.2014.107` | 体外区室选择标准流程 |

### T10 生成式蛋白/酶设计

替代被剔除的 Chem Eur J / JCIM / Biochemistry 等


| 替代文献 | DOI | 作用 |
|---|---|---|
| Nature Methods 2024 · Chroma 生成式蛋白设计模型 | `10.1038/s41592-023-02155-9` | 生成式设计的顶刊代表 |
| Nature Catalysis 2020 · Ensemble-based enzyme design | `10.1038/s41929-020-00529-2` | 计算酶设计的高分替代 |

---


## 六、三条关键结论（如实说明）


### 1. 严格按 IF ≥ 15 过滤后，「保留什么」和「剔除什么」高度分化

| 类别 | 过滤后命运 | 代表 |
|---|---|---|
| **AI/ML 方法与蛋白语言模型** | ✅ 基本保留 | Science（EVOLVEpro）、Nat Commun（主动学习）、Nature Methods |
| **分子诊断的测序读出/纠错** | ✅ 基本保留 | NAR（纠错酶比较、保真度）、Nat Genet（CODEC）、Nat Biotechnol |
| **权威综述与经典源头** | ✅ 保留 | Chem Rev、Chem Soc Rev、NAR（LAMP/PCR 原始文献） |
| **具体酶变体与工程参数** | ❌ 几乎全被剔除 | Bst/Taq/RT 变体的活性倍数、耐抑制阈值、dUTP 兼容性 |
| **冻干配方与常温储运** | ❌ 全部被剔除 | 海藻糖+甘露醇配方、37 ℃ 12 周稳定性 |
| **PCR 抑制机制与临床基质** | ❌ 全部被剔除 | 血液/粪便/腐殖酸阈值、黑色素抑制机理 |

### 2. 有 4 类主题在 IF ≥ 15 期刊中**确实不存在对应文献**

这不是检索不力，而是**学科发表格局**决定的：**配方、工艺参数、变体阈值、诊断应用验证**这类内容天然发表在专业刊（*J Pharm Sci*、*Appl Microbiol Biotechnol*、*Int J Biol Macromol*、*Sci Rep*、*ACS Synth Biol*）。
我们已在高分刊集合（Nature/Science/Cell 系 + Chem Rev + Chem Soc Rev + NAR + Nat Catal 等 20 本）内做了**期刊限定检索**，结果为空：

- **冻干/常温稳定**（T6）：15 本高分刊内检索无一条相关内容
- **Bst/Taq/RT 变体的耐抑制阈值**：高分刊没有"DBS-K431E 耐 50 U/mL 肝素"这类粒度
- **dUTP 兼容聚合酶（防污染 UNG 体系）**：仅见 BMC Biotechnol 等专业刊
- **PCR 抑制剂浓度阈值**：仅见 Forensic Sci Int / Diagnostics 等

### 3. 建议采用「双轨引用」而不是一刀切

| 用途 | 引用来源建议 |
|---|---|
| 讲**趋势、方法、机制**（对外宣讲、立项材料、高层汇报） | 只用 IF ≥ 15（本文件第二节 47 篇） |
| 讲**配方、参数、变体、验证数据**（研发对接、供应商评估、技术方案） | 专业刊必要且合理——行业惯例，客户与评审普遍接受 |
| 两者都要时 | 高分刊引方法，专业刊引数据，**分级标注**（如"方法见 *Science* 2025；参数见 *J Pharm Sci* 2025"） |

> **若坚持只用 IF ≥ 15**：报告可保留 47 篇（AI/ML 方法与读出端），但需删除全部工程参数结论——**那会让报告失去对宝锐最直接的价值**。建议按上表双轨处理。


---


## 七、边界与口径说明（可复核）

| 事项 | 说明 |
|---|---|
| **压线刊** | *Nucleic Acids Research* **15.0**（2025）—— 正好达线，保留；共 19 篇，是保留清单的主力 |
| **边界刊** | *Trends in Biotechnology* **14.9**（2024）/ **16.6**（2025 更新，来源分歧）→ 本表按 14.9 判为剔除；若采用 2025 口径则应保留（仅 1 篇） |
| **已剔除但接近** | *Biotechnology Advances* 14.2、*ACS Catalysis* 13.6、*Nature Chemical Biology* 13.5、*Chem Eng J* 13.2 |
| **无 IF（不可比）** | bioRxiv 预印本、*Nature Chemical Engineering*（2024 新刊，尚无 JCR IF）、*JOSS*（无 IF）、*AAPS Open* / *Frontiers in Sensors* |
| **IF 来源** | Wikipedia 期刊信息框（含年份）+ OpenAlex 两年引用均值交叉验证；边界刊用出版社页面与检索值双源核对 |
| **⚠ 提醒** | 影响因子为 JCR 商业指标，不同年份/来源会有差异；本文件的判定与数值均已标注出处年份，可逐条复核 |